Device for passively removing a target component from blood or lymph of a vertebrate subject

ABSTRACT

Devices, systems, and methods are described herein for controlling or modulating the levels of one or more target components in the blood and/or lymph of a vertebrate subject. Devices and systems are provided that include a body defining at least one lumen configured for fluid flow; at least one controllable flow barrier to fluid flow into the at least one lumen; at least one first reservoir disposed within the body and configured to include one or more bifunctional tags, wherein the one or more bifunctional tags are configured to selectively bind to one or more target components in one or more of blood fluid or lymph fluid of a vertebrate subject; at least one treatment region disposed within the at least one lumen; and at least one second reservoir disposed in the at least one treatment region and configured to include one or more reactive components, wherein the one or more reactive components are configured to sequester the one or more bifunctional tags when bound to the one or more target components.

RELATED APPLICATIONS

For purposes of the USPTO extra-statutory requirements, the presentapplication constitutes a continuation-in-part of U.S. patentapplication Ser. No. TO BE ASSIGNED, entitled DEVICE FOR ACTIVELYREMOVING A TARGET CELL FROM BLOOD OR LYMPH OF A VERTEBRATE SUBJECT,naming RODERICK A. HYDE, MURIEL Y. ISHIKAWA, EDWARD K. Y. JUNG, ROBERTLANGER, ERIC C. LEUTHARDT, NATHAN P. MYHRVOLD, ELIZABETH A. SWEENEY, ANDLOWELL L . WOOD, JR. as inventors, filed 5 Mar. 2010, which is currentlyco-pending, or is an application of which a currently co-pendingapplication is entitled to the benefit of the filing date.

For purposes of the USPTO extra-statutory requirements, the presentapplication constitutes a continuation-in-part of U.S. patentapplication Ser. No. 12/380,400, entitled DEVICE, SYSTEM, AND METHOD FORCONTROLLABLY REDUCING INFLAMMATORY MEDIATORS IN A SUBJECT, namingRODERICK A. HYDE, MURIEL Y. ISHIKAWA, EDWARD K. Y. JUNG, ROBERT LANGER,ERIC C. LEUTHARDT, NATHAN P. MYHRVOLD, ELIZABETH A. SWEENEY, AND LOWELLL. WOOD, JR. as inventors, filed 25 Feb. 2009, which is currentlyco-pending, or is an application of which a currently co-pendingapplication is entitled to the benefit of the filing date.

For purposes of the USPTO extra-statutory requirements, the presentapplication constitutes a continuation-in-part of U.S. patentapplication Ser. No. 12/380,399, entitled DEVICE, SYSTEM, AND METHOD FORCONTROLLABLY REDUCING INFLAMMATORY MEDIATORS IN A SUBJECT, namingRODERICK A. HYDE, MURIEL Y. ISHIKAWA, EDWARD K. Y. JUNG, ROBERT LANGER,ERIC C. LEUTHARDT, NATHAN P. MYHRVOLD, ELIZABETH A. SWEENEY, AND LOWELLL . WOOD, JR. as inventors, filed 25 Feb. 2009, which is currentlyco-pending, or is an application of which a currently co-pendingapplication is entitled to the benefit of the filing date.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is related to and claims the benefit of theearliest available effective filing date(s) from the following listedapplication(s) (the “Related Applications”) (e.g., claims earliestavailable priority dates for other than provisional patent applicationsor claims benefits under 35 USC §119(e) for provisional patentapplications, for any and all parent, grandparent, great-grandparent,etc. applications of the Related Application(s)). All subject matter ofthe Related Applications and of any and all parent, grandparent,great-grandparent, etc. applications of the Related Applications isincorporated herein by reference to the extent such subject matter isnot inconsistent herewith.

The United States Patent Office (USPTO) has published a notice to theeffect that the USPTO's computer programs require that patent applicantsreference both a serial number and indicate whether an application is acontinuation or continuation-in-part. Stephen G. Kunin, Benefit ofPrior-Filed Application, USPTO Official Gazette Mar. 18, 2003, availableat http://www.uspto.gov/web/offices/com/sol/og/2003/week11/patbene.htm.The present Applicant Entity (hereinafter “Applicant”) has providedabove a specific reference to the application(s)from which priority isbeing claimed as recited by statute. Applicant understands that thestatute is unambiguous in its specific reference language and does notrequire either a serial number or any characterization, such as“continuation” or “continuation-in-part,” for claiming priority to U.S.patent applications. Notwithstanding the foregoing, Applicantunderstands that the USPTO's computer programs have certain data entryrequirements, and hence Applicant is designating the present applicationas a continuation-in-part of its parent applications as set forth above,but expressly points out that such designations are not to be construedin any way as any type of commentary and/or admission as to whether ornot the present application contains any new matter in addition to thematter of its parent application(s).

SUMMARY

Devices, systems, and methods are disclosed herein for controlling ormodulating the levels of one or more target components in the bloodfluid and/or lymph fluid of a vertebrate subject. The implantable deviceor system can include a body defining at least one lumen configured forfluid flow. The device or system can further include at least one firstreservoir disposed within the at least one lumen and configured toinclude one or more bifunctional tags, wherein the one or morebifunctional tags are configured to selectively bind to one or moretarget components in one or more of blood fluid or lymph fluid of avertebrate subject, and at least one treatment region disposed withinthe at least one lumen. The device or system can further include atleast one second reservoir disposed in the at least one treatment regionand configured to include one or more reactive components, wherein theone or more reactive components are configured to sequester the one ormore bifunctional tags when bound to the one or more target components.

The device or system is useful in a method for treating a disease orcondition mediated by or indicated by the one or more target components.The one or more target components can include, but are not limited to,cellular components (e.g., blood cells, cancer cells, pathogens),non-cellular components (e.g., proteins, lipids, sugars, carbohydrates,small molecules), or combinations thereof. Examples of diseases,conditions, or infections include, but are not limited to, acute andchronic inflammatory diseases (e.g., sepsis, multiple organ dysfunctionsyndrome, autoimmune disease, asthma, rhinitis, rheumatoid arthritis),cardiovascular disease, gastrointestinal disease, neoplastic disease,metabolic disease, bacterial infection (e.g., Staphylococcusbacteremia), viral infection (e.g., acquired immunodeficiency syndrome,hepatitis), parasite infection (e.g., malaria), or chemical orbiological agent exposure (e.g., drug overdose, environmental toxin).

An implantable device is described that includes a body defining atleast one lumen configured for fluid flow; at least one first reservoirdisposed within the at least one lumen and configured to include one ormore bifunctional tags, wherein the one or more bifunctional tags areconfigured to selectively bind to one or more target components in oneor more of blood fluid or lymph fluid of a vertebrate subject; at leastone treatment region disposed within the at least one lumen; and atleast one second reservoir disposed in the at least one treatment regionand configured to include one or more reactive components, wherein theone or more reactive components are configured to sequester the one ormore bifunctional tags when bound to the one or more target components.The one or more reactive components can be configured to have anincreased affinity for the one or more bifunctional tags bound to theone or more target components compared to an affinity for the one ormore bifunctional tags unbound to the one or more reactive components.The one or more reactive components can be configured to modulate aphysiological effect of the one or more target components. The one ormore bifunctional tags can be configured to enter a circulatory systemof the mammalian subject at a site different from a site of the one ormore reactive components. In an aspect, the one or more bifunctionaltags can include, but is not limited to, one or more of a recognitionelement, recognition molecule, antibody, integrin, selectin, lectin,mimetic polymer, affibody, a label, or virus-like particle. The labelcan include, but is not limited to, one or more of a QDOT, ananoparticle, a fluorescent molecule, a magnetic particle, a contrastagent, or a radioisotope. The one or more bifunctional tags can includeone or more bifunctional antibodies. The one or more bifunctionalantibodies is configured to bind to one or more of the target componentand the reactive component. The one or more target components caninclude one or more of circulating target cells or circulating targetemboli. The one or more target components can include, but is notlimited to, one or more of tumor cells, emboli, misfolded proteins,aggregated proteins, antibodies, autoimmune antibodies, infectiousagents, or infected cells. The one or more target components caninclude, but is not limited to, cancer cells, pre-cancer cells,autoimmune-related cells, B cells, T cells, phagocytes, platelets,lipoproteins, parasites, viruses, bacteria, fungi, or infected cells.The one or more reactive components can be configured to attach to theat least one lumen. The device can further include two or more parallellumen configured to receive the one or more target components. In anaspect, a diameter of each of the two or more lumen can be approximatelyless than two cell diameters. In an aspect, a diameter of each of thetwo or more lumen can be approximately less than 10 μm.

The device including the at least one second reservoir disposed in theat least one treatment region and configured to include one or morereactive components, wherein the reactive components include two or morebinding agents. The two or more binding agents are configured tosequester the one or more bifunctional tags when bound to the one ormore target components. One or more binding agents can include bindingagents, e.g., selectins or integrins, to slow passage of the targetcomponent through the treatment region. Other of the one or more bindingagents can include sequestering agents, e.g., antibodies or mimetics, tosequester the one or more target component within the treatment region.

The one or more reactive components can be configured to alter, arrest,or destroy the one or more target cells. The one or more reactivecomponents can be configured to produce necrosis or apoptosis in one ormore target cells. The one or more reactive components can be configuredto alter, arrest, or destroy the one or more target components. The oneor more reactive components can be configured to be placed relative to atumor or an organ in the mammalian subject. The one or more reactivecomponents can include, but is not limited to, one or more of anadhesion molecule, antibody, binding mimetic, polymer, lectin, integrin,or selectin. The one or more reactive components can include, but is notlimited to, one or more of a denaturing agent, degradative agent, orbinding agent. The one or more reactive components can include, but isnot limited to, a cytotoxic agent, a cytostatic agent, a programmed celldeath-inducing agent, a chemotherapeutic agent, or an antibody-toxinagent. A programmed cell death-inducing agent can include an agent orcompound that induces programmed cell death in eukaryotic or prokaryoticcells, or an agent or compound that induces apoptosis in eukaryoticcells. The one or more binding agents can include, but is not limitedto, one or more of antibodies, receptors, or cognates configured to bindto one or more target components. The one or more binding agents caninclude, but is not limited to, one or more of lectin, binding protein,catalytic antibody, catalytic aptamer, protease conjugate, orphotoactivatable conjugate. The device can further include one or moreenergy sources configured to supply energy to the at least one treatmentregion.

The device can further include one or more sensor configured to measurea physiological condition proximate to the device. The one or moresensor can be configured to detect one or more of unbound bifunctionaltags or bifunctional tags bound to target components in the one or moreof blood fluid or lymph fluid of the vertebrate subject. The one or moresensor can be configured to detect sequestration by the at least onereactive component. The device can further include a transmitter toreport data from the one or more sensor. The one or more sensor can beconfigured to report to an outside source or to a computing device. Theone or more sensor can be configured to function in, or proximal to, theone or more blood vessel or lymph vessel. The one or more sensor can beexternal to the at least one lumen. The one or more sensor can beinternal to the at least one lumen. The device can further include atleast one controller in communication with the one or more sensor. Theat least one controller can include a processor. The at least onecontroller in communication with the one or more sensor can beconfigured to control the at least one controllable flow barrier to theat least one lumen.

The one or more sensor can be configured to detect the one or moretarget components and configured to communicate with the at least onecontroller to release the one or more bifunctional tags in response tothe one or more detected target components. The one or more sensor canbe configured to detect the one or more bifunctional tags complexed withthe one or more target components and is configured to communicate withthe controller to activate the one or more reactive components inresponse to the complex of the one or more bifunctional tags to the oneor more target components. The one or more sensor can be configured todetect the one or more bifunctional tags complexed with the one or moretarget components and is configured to communicate with the controllerto divert flow of the one or more of blood fluid or lymph fluid to theat least one lumen of the device. The one or more sensor can beconfigured to detect the one or more bifunctional tags complexed withthe one or more target components and is configured to communicate withthe controller to divert flow of the one or more of blood fluid or lymphfluid away from the at least one lumen of the device. The one or moresensor and the at least one controller can be configured to achieve atarget level of the one or more target components in the vertebratesubject. The one or more sensor and the at least one controller can beconfigured to control the at least one controllable flow barrier, toactivate the one or more reactive components, to release the one or morebifunctional tags, or to activate one or more energy sources. The one ormore sensor and the at least one controller can be configured to controllevels of the detected one or more target components to limit adeviation from the target level. In an aspect, the deviation can bedetermined by a weighted least squares fit. The target level can includea desired concentration of the one or more target components in the oneor more of blood fluid or lymph fluid. The target level can include adesired range of concentrations of the one or more target components inthe one or more of blood fluid or lymph fluid. The target level caninclude a desired ratio of concentrations of two or more targetcomponents in the one or more of blood fluid or lymph fluid. The targetlevel can include a desired ratio of levels of two or more targetcomponents in the one or more of blood fluid or lymph fluid.

The one or more sensor can include, but is not limited to, a biosensor,chemical sensor, physical sensor, or optical sensor. The one or moresensor can include one or more target recognition elements. The one ormore target recognition elements can include, but is not limited to, oneor more of an aptamer, antibody, receptor, affibody, mimic, nucleicacid, or synthetic compound. The one or more sensor can include, but isnot limited to, one or more of a recognition-based substrate, anaptamer-based substrate, an antibody-based substrate, surface plasmonresonance, genetically modified cells, or genetically modified cellswith receptor-linked signaling. The genetically modified cells caninclude receptor-linked signaling by fluorogen-activating proteins. Theone or more sensor can be configured to target the device to a sitehaving an elevated level of the one or more target components. The oneor more sensor can be configured to detect one or more of labeledbifunctional tags, bifunctional tags bound to the one or more targetcomponents, bifunctional tags bound to the one or more target componentsand to the one or more reactive components, or the one or more targetcomponents sequestered by the one or more bifunctional tags and the oneor more reactive components. The device can be configured to report toan outside source or to a computing device, wherein the device isconfigured to report sequestration of the one or more target components.

The at least one first reservoir can be configured to provide one ormore bifunctional tags is responsive to the controller. The at least onesecond reservoir can be configured to provide one or more reactivecomponents is responsive to the controller. The device can furtherinclude at least one controllable flow barrier to fluid flow into the atleast one lumen. The at least one controllable flow barrier can beconfigured to be at least partially open. In an aspect, a portion of theat least one lumen including the at least one first reservoir can bephysically separated from a portion of the at least one lumen includingthe at least one second reservoir.

The one or more sensor can be configured to detect one or more ofT-lymphocytes, B-lymphocytes, antibodies, pre-cancer cells, cancercells, inflammatory cells, infected cells, bacteria, parasites, fungi,viruses, platelets, phagocytes, or lipoproteins. The one or more sensorcan be configured to detect one or more of anaphylatoxins, cytokines,chemokines, leukotrienes, prostaglandins, complements, coagulationfactors, or proinflammatory cytokines. The one or more sensor can beconfigured to detect one or more of body temperature, vital signs,edema, oxygen level, or pathogen/toxin level of the subject. The one ormore sensor can be configured to detect one or more of TNF-α, IL-1,IL-1β, IL-6, IL-8, IL-10, IL-12, LPB, IFN-γ, LIF, MIF, MIP-1, MCP-1,C3-a, or C5-a. The one or more sensor can be configured to detect one ormore of exotoxins, endotoxins, or lipopolysaccharide.

The one or more second reservoirs can include a matrix configured topresent one or more reactive components. The one or more secondreservoirs can include a matrix configured to present one or morereactive components. The one or more binding agents can include one ormore target recognition elements. The one or more target recognitionelements can include, but is not limited to, one or more of aptamer,antibodies, receptors, affibody, mimic, nucleic acid, syntheticcompound, or cognates configured to bind to at least one of the one ormore target components. The binding agent can include one or more of aspecific binding ligand or a hydrophobic surface. The matrix caninclude, but is not limited to, one or more of beads, cells, vesicles,filters, hydrogel polymers, microparticles, nanoparticles, adsorbent,absorbent, or synthetic polymers. The specific binding ligand or thehydrophobic surface can include, but is not limited to, one or more ofnucleic acid aptamers, peptide aptamers, molecular imprinting polymer,antibodies or fragments thereof, high affinity mimetics, syntheticbinding molecules, or receptor binding molecules. The matrix caninclude, but is not limited to, one or more of a lectin, bindingprotein, receptor, antibody, catalytic antibody, catalytic aptamer,protease conjugate, or photoactivatable conjugate.

The energy source can include, but is not limited to, acoustic energy orelectronic energy. The energy source can include ultrasound. In anaspect, the energy source can include high-intensity focused ultrasound.The energy source can include, but is not limited to, at least one ofmicrowave irradiation, gamma irradiation, electromagnetic irradiation,heat, electron beam irradiation, vibrational/frequency irradiation, oratmospheric pressure glow discharge. The vibrational/frequencyirradiation can include a set of differing energy inputs specificallydirected to the one or more target components, wherein the set ofdiffering energy inputs selectively resonates a plurality of resonantstructures in the one or more target components, and wherein theresonance controllably alters or reduces the activity of the one or moretarget components in the one or more of blood fluid or lymph fluid ofthe vertebrate subject. In an aspect, the one or more target componentscan be modified with a functional group configured to be responsive tothe set of differing energy inputs. The energy source can be coupled toone or more sensor configured to selectively direct energy to the targetcomponent. The one or more denaturing agents can include, but is notlimited to, at least of an acid, base, solvent, detergent, cross-linkingagent, chaotropic agent, disulfide bond reducer, enzyme, drug, cell, orradical ion. The one or more degradative agents can include, but is notlimited to, at least one of an enzyme, coenzyme, enzyme complex,catalytic antibody, proteasome, strong acid, strong base, radical,photoactivatable agent, drug, cell, or radical ion. In an aspect, thecatalytic antibody can generate a radical ion. The one or more secondreservoirs can include a source for producing the one or more reactivecomponents. The source can include at least one reservoir and at leastone producer. The at least one producer can include at least oneencapsulated cell. The at least one encapsulated cell can produce theone or more reactive components. The at least one encapsulated cell caninclude at least one genetically-engineered cell. The at least oneencapsulated cell can include, but is not limited to, at least one of amammalian cell, bacterial cell, yeast cell, plant cell, insect cell,artificial cell, or enucleated cell. The at least one encapsulated cellcan include, but is not limited to, one or more of a myeloid cell,lymphocyte, or precursor thereof. The at least one encapsulated cell caninclude, but is not limited to, one or more of a T-lymphocyte,B-lymphocyte, macrophage, monocyte, neutrophil, or NK cell. The at leastone producer can include, but is not limited to, a protein, lipidmicelle, liposome, synthetic polymer, or a combination thereof. The atleast one producer can include a catalytic antibody. In an aspect, thecatalytic antibody can include a radical ion generator.

In an aspect, the device can be intracorporeal. The device can be mobilewithin the blood vessel or lymph vessel. The device can be configured tobe implanted. The device can include, but is not limited to, a stent,bypass implant, nanostructure or microstructure. The device can beconfigured to be implanted relative to an organ or tissue in thesubject. The device can be at least partially extracorporeal. The devicecan include, but is not limited to, a dialysis device, hemoperfusiondevice, apheresis device, intravenous device, shunt device, or patchdevice.

A method for treating an inflammatory condition or inflammatory diseasein a vertebrate subject is described that includes providing animplantable device including a body defining at least one lumenconfigured for fluid flow; at least one first reservoir disposed withinthe at least one lumen and configured to include one or morebifunctional tags, wherein the one or more bifunctional tags areconfigured to selectively bind to one or more target components in oneor more of blood fluid or lymph fluid of a vertebrate subject; at leastone treatment region disposed within the at least one lumen; and atleast one second reservoir disposed in the at least one treatment regionand configured to include one or more reactive components, wherein theone or more reactive components are configured to sequester the one ormore bifunctional tags when bound to the one or more target components.The one or more reactive components can be configured to have anincreased affinity for the one or more bifunctional tags bound to theone or more target components compared to an affinity for the one ormore bifunctional tags unbound to the one or more reactive components.The one or more reactive components can be configured to modulate aphysiological effect of the one or more target components. The one ormore bifunctional tags can be configured to enter a circulatory systemof the mammalian subject at a site different from a site of the one ormore reactive components. In an aspect, the one or more bifunctionaltags can include, but is not limited to, one or more of a recognitionelement, recognition molecule, antibody, integrin, selectin, lectin,mimetic polymer, affibody, a label, or virus-like particle. The labelcan include, but is not limited to, one or more of a QDOT, ananoparticle, a fluorescent molecule, a magnetic particle, a contrastagent, or a radioisotope. The one or more bifunctional tags can includeone or more bifunctional antibodies. The one or more bifunctionalantibodies is configured to bind to one or more of the target componentand the reactive component. The one or more target components caninclude one or more of circulating target cells or circulating targetemboli. The one or more target components can include, but is notlimited to, one or more of tumor cells, emboli, misfolded proteins,aggregated proteins, antibodies, autoimmune antibodies, infectiousagents, or infected cells. The one or more target components caninclude, but is not limited to, cancer cells, pre-cancer cells,autoimmune-related cells, B cells, T cells, phagocytes, platelets,lipoproteins, parasites, viruses, bacteria, fungi, or infected cells.The one or more reactive components can be configured to attach to theat least one lumen.

The method can further include providing two or more parallel lumenconfigured to receive the one or more target components. In an aspect, adiameter of each of the two or more lumen can be approximately less thantwo cell diameters. In an aspect, a diameter of each of the two or morelumen can be approximately less than 10 μm.

The one or more reactive components can be configured to alter, arrest,or destroy the one or more target cells. The one or more reactivecomponents can be configured to produce necrosis or apoptosis in one ormore target cells. The one or more reactive components can be configuredto alter, arrest, or destroy the one or more target components. The oneor more reactive components can be configured to be placed relative to atumor or an organ in the mammalian subject. The one or more reactivecomponents can include, but is not limited to, one or more of anadhesion molecule, antibody, binding mimetic, polymer, lectin, integrin,or selectin. The one or more reactive components can include, but is notlimited to, one or more of a denaturing agent, degradative agent, orbinding agent. The one or more reactive components can include, but isnot limited to, a cytotoxic agent, a cytostatic agent, a programmed celldeath-inducing agent, a chemotherapeutic agent, or an antibody-toxinagent. The one or more binding agents can include, but is not limitedto, one or more of antibodies, receptors, or cognates configured to bindto one or more target components. The one or more binding agents caninclude, but is not limited to, one or more of lectin, binding protein,catalytic antibody, catalytic aptamer, protease conjugate, orphotoactivatable conjugate. The method can further include providing oneor more energy sources configured to supply energy to the at least onetreatment region.

The method can further include providing one or more sensors configuredto measure a physiological condition proximate to the device. The one ormore sensor can be configured to detect one or more of unboundbifunctional tags or bifunctional tags bound to target components in theone or more of blood fluid or lymph fluid of the vertebrate subject. Theone or more sensor can be configured to detect sequestration by the atleast one reactive component. The method can further include providing atransmitter to report data from the one or more sensor. The one or moresensor can be configured to report to an outside source or to acomputing device. The one or more sensor can be configured to functionin, or proximal to, the one or more blood vessel or lymph vessel. Theone or more sensor can be external to the at least one lumen. The one ormore sensor can be internal to the at least one lumen. The method canfurther include providing at least one controller in communication withthe one or more sensor. The at least one controller can include aprocessor. The method can further include providing at least onecontrollable flow barrier to fluid flow into the at least one lumen. Theat least one controllable flow barrier can be configured to be at leastpartially open. The at least one controller in communication with theone or more sensor can be configured to control the at least onecontrollable flow barrier to the at least one lumen.

The one or more sensor can be configured to detect the one or moretarget components and configured to communicate with the at least onecontroller to release the one or more bifunctional tags in response tothe one or more detected target components. The one or more sensor canbe configured to detect the one or more bifunctional tags complexed withthe one or more target components and is configured to communicate withthe controller to activate the one or more reactive components inresponse to the complex of the one or more bifunctional tags to the oneor more target components. The one or more sensor can be configured todetect the one or more bifunctional tags complexed with the one or moretarget components and is configured to communicate with the controllerto divert flow of the one or more of blood fluid or lymph fluid to theat least one lumen of the device. The one or more sensor can beconfigured to detect the one or more bifunctional tags complexed withthe one or more target components and is configured to communicate withthe controller to divert flow of the one or more of blood fluid or lymphfluid away from the at least one lumen of the device. The one or moresensor and the at least one controller can be configured to achieve atarget level of the one or more target components in the vertebratesubject. The one or more sensor and the at least one controller can beconfigured to control the at least one controllable flow barrier, toactivate the one or more reactive components, to release the one or morebifunctional tags, or to activate one or more energy sources. The one ormore sensor and the at least one controller can be configured to controllevels of the detected one or more target components to limit adeviation from the target level. In an aspect, the deviation can bedetermined by a weighted least squares fit. The target level can includea desired concentration of the one or more target components in the oneor more of blood fluid or lymph fluid. The target level can include adesired range of concentrations of the one or more target components inthe one or more of blood fluid or lymph fluid. The target level caninclude a desired ratio of concentrations of two or more targetcomponents in the one or more of blood fluid or lymph fluid. The targetlevel can include a desired ratio of levels of two or more targetcomponents in the one or more of blood fluid or lymph fluid.

The one or more sensor can include, but is not limited to, a biosensor,chemical sensor, physical sensor, or optical sensor. The one or moresensor can include one or more target recognition elements. The one ormore target recognition elements can include, but is not limited to, oneor more of an aptamer, antibody, receptor, affibody, mimic, nucleicacid, or synthetic compound. The one or more sensor can include, but isnot limited to, one or more of a recognition-based substrate, anaptamer-based substrate, an antibody-based substrate, surface plasmonresonance, genetically modified cells, or genetically modified cellswith receptor-linked signaling. The genetically modified cells caninclude receptor-linked signaling by fluorogen-activating proteins. Theone or more sensor can be configured to target the device to a sitehaving an elevated level of the one or more target components. The oneor more sensor can be configured to detect one or more of labeledbifunctional tags, bifunctional tags bound to the one or more targetcomponents, bifunctional tags bound to the one or more target componentsand to the one or more reactive components, or the one or more targetcomponents sequestered by the one or more bifunctional tags and the oneor more reactive components. The device can be configured to report toan outside source or to a computing device, wherein the device isconfigured to report sequestration of the one or more target components.

The at least one first reservoir can be configured to provide one ormore bifunctional tags is responsive to the controller. The at least onesecond reservoir can be configured to provide one or more reactivecomponents is responsive to the controller. In an aspect, a portion ofthe at least one lumen including the at least one first reservoir can bephysically separated from a portion of the at least one lumen includingthe at least one second reservoir.

The one or more sensor can be configured to detect one or more ofT-lymphocytes, B-lymphocytes, antibodies, pre-cancer cells, cancercells, inflammatory cells, infected cells, bacteria, parasites, fungi,viruses, platelets, phagocytes, or lipoproteins. The one or more sensorcan be configured to detect one or more of anaphylatoxins, cytokines,chemokines, leukotrienes, prostaglandins, complements, coagulationfactors, or proinflammatory cytokines. The one or more sensor can beconfigured to detect one or more of body temperature, vital signs,edema, oxygen level, or pathogen/toxin level of the subject. The one ormore sensor can be configured to detect one or more of TNF-α, IL-1,IL-1β, IL-6, IL-8, IL-10, IL-12, LPB, IFN-γ, LIF, MIF, MIP-1, MCP-1,C3-a, or C5-a. The one or more sensor can be configured to detect one ormore of exotoxins, endotoxins, or lipopolysaccharide.

The one or more second reservoirs can include a matrix configured topresent one or more reactive components. The one or more secondreservoirs can include a matrix configured to present one or morereactive components. The one or more binding agents can include one ormore target recognition elements. The one or more target recognitionelements can include, but is not limited to, one or more of aptamer,antibodies, receptors, affibody, mimic, nucleic acid, syntheticcompound, or cognates configured to bind to at least one of the one ormore target components. The binding agent can include one or more of aspecific binding ligand or a hydrophobic surface. The matrix caninclude, but is not limited to, one or more of beads, cells, vesicles,filters, hydrogel polymers, microparticles, nanoparticles, adsorbent,absorbent, or synthetic polymers. The specific binding ligand or thehydrophobic surface can include, but is not limited to, one or more ofnucleic acid aptamers, peptide aptamers, molecular imprinting polymer,antibodies or fragments thereof, high affinity mimetics, syntheticbinding molecules, or receptor binding molecules. The matrix caninclude, but is not limited to, one or more of a lectin, bindingprotein, receptor, antibody, catalytic antibody, catalytic aptamer,protease conjugate, or photoactivatable conjugate.

The energy source can include, but is not to, acoustic energy orelectronic energy. The energy source can include ultrasound. In anaspect, the energy source can include high-intensity focused ultrasound.The energy source can include, but is not limited to, at least one ofmicrowave irradiation, gamma irradiation, electromagnetic irradiation,heat, electron beam irradiation, vibrational/frequency irradiation, oratmospheric pressure glow discharge. The vibrational/frequencyirradiation can include a set of differing energy inputs specificallydirected to the one or more target components, wherein the set ofdiffering energy inputs selectively resonates a plurality of resonantstructures in the one or more target components, and wherein theresonance controllably alters or reduces the activity of the one or moretarget components in the one or more of blood fluid or lymph fluid ofthe vertebrate subject. In an aspect, the one or more target componentscan be modified with a functional group configured to be responsive tothe set of differing energy inputs. The energy source can be coupled toone or more sensor configured to selectively direct energy to the targetcomponent. The one or more denaturing agents can include, but is notlimited to, at least of an acid, base, solvent, detergent, cross-linkingagent, chaotropic agent, disulfide bond reducer, enzyme, drug, cell, orradical ion. The one or more degradative agents can include, but is notlimited to, at least one of an enzyme, coenzyme, enzyme complex,catalytic antibody, proteasome, strong acid, strong base, radical,photoactivatable agent, drug, cell, or radical ion. In an aspect, thecatalytic antibody can generate a radical ion. The one or more secondreservoirs can include a source for producing the one or more reactivecomponents. The source can include at least one reservoir and at leastone producer. The at least one producer can include at least oneencapsulated cell. The at least one encapsulated cell can produce theone or more reactive components. The at least one encapsulated cell caninclude at least one genetically-engineered cell. The at least oneencapsulated cell can include, but is not limited to, at least one of amammalian cell, bacterial cell, yeast cell, plant cell, insect cell,artificial cell, or enucleated cell. The at least one encapsulated cellcan include, but is not limited to, one or more of a myeloid cell,lymphocyte, or precursor thereof. The at least one encapsulated cell caninclude, but is not limited to, one or more of a T-lymphocyte,B-lymphocyte, macrophage, monocyte, neutrophil, or NK cell. The at leastone producer can include, but is not limited to, a protein, lipidmicelle, liposome, synthetic polymer, or a combination thereof. The atleast one producer can include a catalytic antibody. In an aspect, thecatalytic antibody can include a radical ion generator.

A method for modulating an inflammatory condition or inflammatorydisease in a vertebrate subject is described that includes providing animplantable device including a body defining at least one lumenconfigured for fluid flow; at least one controllable flow barrier tofluid flow into the at least one lumen; at least one first reservoirdisposed within the body and configured to include one or morebifunctional tags, wherein the one or more bifunctional tags areconfigured to selectively bind to one or more target components in oneor more of blood fluid or lymph fluid of a vertebrate subject; at leastone treatment region disposed within the at least one lumen; and atleast one second reservoir disposed in the at least one treatment regionand configured to include one or more reactive components, wherein theone or more reactive components are configured to sequester the one ormore bifunctional tags when bound to the one or more target components.The one or more reactive components can have an increased affinity forthe one or more bifunctional tags bound to the one or more targetcomponents compared to an affinity for the one or more bifunctional tagsunbound to the one or more reactive components. The one or more reactivecomponents can be configured to modulate a physiological effect of theone or more target components. The one or more reactive components caninclude, but is not limited to, a one or more of a denaturing agent,degradative agent or binding agent.

A method for treating an infectious disease or infectious condition in avertebrate subject is described that includes providing an implantabledevice including a body defining at least one lumen configured for fluidflow; at least one controllable flow barrier to fluid flow into the atleast one lumen; at least one first reservoir disposed within the bodyand configured to include one or more bifunctional tags, wherein the oneor more bifunctional tags are configured to selectively bind to one ormore target components in one or more of blood fluid or lymph fluid of avertebrate subject; at least one treatment region disposed within the atleast one lumen; and at least one second reservoir disposed in the atleast one treatment region and configured to include one or morereactive components, wherein the one or more reactive components areconfigured to sequester the one or more bifunctional tags when bound tothe one or more target components. The one or more reactive componentscan have an increased affinity for the one or more bifunctional tagsbound to the one or more target components compared to an affinity forthe one or more bifunctional tags unbound to the one or more reactivecomponents. The one or more reactive components can be configured tomodulate a physiological effect of the one or more target components.The one or more reactive components can include, but is not limited to,a one or more of a denaturing agent, degradative agent or binding agent.

A method for modulating an infectious disease or infectious condition ina vertebrate subject is described that includes providing an implantabledevice including a body defining at least one lumen configured for fluidflow; at least one controllable flow barrier to fluid flow into the atleast one lumen; at least one first reservoir disposed within the bodyand configured to include one or more bifunctional tags, wherein the oneor more bifunctional tags are configured to selectively bind to one ormore target components in one or more of blood fluid or lymph fluid of avertebrate subject; at least one treatment region disposed within the atleast one lumen; and at least one second reservoir disposed in the atleast one treatment region and configured to include one or morereactive components, wherein the one or more reactive components areconfigured to sequester the one or more bifunctional tags when bound tothe one or more target components. The one or more reactive componentscan have an increased affinity for the one or more bifunctional tagsbound to the one or more target components compared to an affinity forthe one or more bifunctional tags unbound to the one or more reactivecomponents. The one or more reactive components can be configured tomodulate a physiological effect of the one or more target components.The one or more reactive components can include, but is not limited to,a one or more of a denaturing agent, degradative agent or binding agent.

A method for treating a neoplastic disease or neoplastic condition in avertebrate subject is described that includes providing an implantabledevice including a body defining at least one lumen configured for fluidflow; at least one controllable flow barrier to fluid flow into the atleast one lumen; at least one first reservoir disposed within the bodyand configured to include one or more bifunctional tags, wherein the oneor more bifunctional tags are configured to selectively bind to one ormore target components in one or more of blood fluid or lymph fluid of avertebrate subject; at least one treatment region disposed within the atleast one lumen; and at least one second reservoir disposed in the atleast one treatment region and configured to include one or morereactive components, wherein the one or more reactive components areconfigured to sequester the one or more bifunctional tags when bound tothe one or more target components.

A method for modulating a neoplastic disease or neoplastic condition ina vertebrate subject is described that includes providing an implantabledevice including a body defining at least one lumen configured for fluidflow; at least one controllable flow barrier to fluid flow into the atleast one lumen; at least one first reservoir disposed within the bodyand configured to include one or more bifunctional tags, wherein the oneor more bifunctional tags are configured to selectively bind to one ormore target components in one or more of blood fluid or lymph fluid of avertebrate subject; at least one treatment region disposed within the atleast one lumen; and at least one second reservoir disposed in the atleast one treatment region and configured to include one or morereactive components, wherein the one or more reactive components areconfigured to sequester the one or more bifunctional tags when bound tothe one or more target components. The one or more reactive componentscan have an increased affinity for the one or more bifunctional tagsbound to the one or more target components compared to an affinity forthe one or more bifunctional tags unbound to the one or more reactivecomponents. The one or more reactive components can be configured tomodulate a physiological effect of the one or more target components.The one or more reactive components can include, but is not limited to,a one or more of a denaturing agent, degradative agent or binding agent.

A system is described that includes an implantable device including abody defining at least one lumen configured for fluid flow; at least onefirst reservoir disposed within the body and configured to include oneor more bifunctional tags, wherein the one or more bifunctional tags areconfigured to selectively bind to one or more target components in oneor more of blood fluid or lymph fluid of a vertebrate subject; at leastone treatment region disposed within the at least one lumen; and atleast one second reservoir disposed in the at least one treatment regionand configured to include one or more reactive components, wherein theone or more reactive components are configured to sequester the one ormore bifunctional tags when bound to the one or more target components.The one or more reactive components can have an increased affinity forthe one or more bifunctional tags bound to the one or more targetcomponents compared to an affinity for the one or more bifunctional tagsunbound to the one or more reactive components. The one or more reactivecomponents can be configured to modulate a physiological effect of theone or more target components. The one or more reactive components caninclude, but is not limited to, a one or more of a denaturing agent,degradative agent or binding agent.

A device is described that includes a system including a signal-bearingmedium including one or more instructions for treatment of a subjectthrough an implantable device including a body defining at least onelumen configured for fluid flow; at least one controllable flow barrierto fluid flow into the at least one lumen; at least one first reservoirdisposed within the body and configured to include one or morebifunctional tags, wherein the one or more bifunctional tags areconfigured to selectively bind to one or more target components in oneor more of blood fluid or lymph fluid of a vertebrate subject; at leastone treatment region disposed within the at least one lumen; and atleast one second reservoir disposed in the at least one treatment regionand configured to include one or more reactive components, wherein theone or more reactive components are configured to sequester the one ormore bifunctional tags when bound to the one or more target components;and one or more instructions for receiving data including data fordelivering one or more reactive components configured to sequester theone or more bifunctional tags when bound to the one or more targetcomponents.

A system is described that includes at least one computer programincluded on a computer-readable medium for use with at least onecomputer system wherein the computer program includes a plurality ofinstructions including, one or more instructions for determining atleast one treatment of one or more of blood fluid or lymph fluid of avertebrate subject through an implantable device including a bodydefining at least one lumen configured for fluid flow; at least onecontrollable flow barrier to fluid flow into the at least one lumen; atleast one first reservoir disposed within the body and configured toinclude one or more bifunctional tags, wherein the one or morebifunctional tags are configured to selectively bind to one or moretarget components in the one or more of blood fluid or lymph fluid of avertebrate subject; at least one treatment region disposed within the atleast one lumen; and at least one second reservoir disposed in the atleast one treatment region and configured to include one or morereactive components, wherein the one or more reactive components areconfigured to sequester the one or more bifunctional tags when bound tothe one or more target components; and one or more instructions forreceiving data including data for delivering one or more reactivecomponents configured to sequester the one or more bifunctional tagswhen bound to the one or more target components.

The foregoing summary is illustrative only and is not intended to be inany way limiting. In addition to the illustrative aspects, embodiments,and features described above, further aspects, embodiments, and featureswill become apparent by reference to the drawings and the followingdetailed description.

BRIEF DESCRIPTION OF THE FIGURES

FIGS. 1A and 1B depict a diagrammatic view of an aspect of an embodimentof a device.

FIG. 2 depicts a diagrammatic view of an aspect of an embodiment of adevice.

FIG. 3 depicts a diagrammatic view of an aspect of an embodiment of adevice.

DETAILED DESCRIPTION

In the following detailed description, reference is made to theaccompanying drawings, which form a part hereof. In the drawings,similar symbols typically identify similar components, unless contextdictates otherwise. The illustrative embodiments described in thedetailed description, drawings, and claims are not meant to be limiting.Other embodiments may be utilized, and other changes may be made,without departing from the spirit or scope of the subject matterpresented here.

This document uses formal outline headings for clarity of presentation.However, it is to be understood that the outline headings are forpresentation purposes, and that different types of subject matter may bediscussed throughout the application (e.g., method(s) may be describedunder composition heading(s) and/or kit headings, arid/or descriptionsof single topics may span two or more topic headings). Hence, the use ofthe formal outline headings is not intended to be in any way limiting.Devices, systems, and methods are disclosed herein for controlling ormodulating the levels of one or more target components in the bloodfluid and/or lymph fluid of a vertebrate subject. The implantable deviceor system can include a body defining at least one lumen configured forfluid flow. The device or system can further include at least one firstreservoir disposed within the at least one lumen and configured toinclude one or more bifunctional tags, wherein the one or morebifunctional tags are configured to selectively bind to one or moretarget components in one or more of blood fluid or lymph fluid of avertebrate subject, and at least one treatment region disposed withinthe at least one lumen. The device or system can further include atleast one second reservoir disposed in the at least one treatment regionand configured to include one or more reactive components, wherein theone or more reactive components are configured to sequester the one ormore bifunctional tags when bound to the one or more target components.

The device or system is useful in a method for treating a disease orcondition mediated by or indicated by the one or more target components.The one or more target components can include, but are not limited to,cellular components (e.g., blood cells, cancer cells, pathogens),non-cellular components (e.g., proteins, lipids, sugars, carbohydrates,small molecules), or combinations thereof. Examples of diseases,conditions, or infections include, but are not limited to, acute andchronic inflammatory diseases (e.g., sepsis, multiple organ dysfunctionsyndrome, autoimmune disease, asthma, rhinitis, rheumatoid arthritis),cardiovascular disease, gastrointestinal disease, neoplastic disease,metabolic disease, bacterial infection (e.g., Staphylococcusbacteremia), viral infection (e.g., acquired immunodeficiency syndrome,hepatitis), parasite infection (e.g., malaria), or chemical orbiological agent exposure (e.g., drug overdose, environmental toxin).

An implantable device is described that includes a body defining atleast one lumen configured for fluid flow; at least one first reservoirdisposed within the at least one lumen and configured to include one ormore bifunctional tags, wherein the one or more bifunctional tags areconfigured to selectively bind to one or more target components in oneor more of blood fluid or lymph fluid of a vertebrate subject; at leastone treatment region disposed within the at least one lumen; and atleast one second reservoir disposed in the at least one treatment regionand configured to include one or more reactive components, wherein theone or more reactive components are configured to sequester the one ormore bifunctional tags when bound to the one or more target components.The one or more reactive components can be configured to have anincreased affinity for the one or more bifunctional tags bound to theone or more target components compared to an affinity for the one ormore bifunctional tags unbound to the one or more reactive components.The one or more reactive components can be configured to modulate aphysiological effect of the one or more target components. In an aspect,the one or more bifunctional tags can include, but is not limited to,one or more of a recognition element, recognition molecule, antibody,integrin, selectin, lectin, mimetic polymer, affibody, a label, orvirus-like particle. The label can include, but is not limited to, oneor more of a QDOT, a nanoparticle, a fluorescent molecule, a magneticparticle, a contrast agent, or a radioisotope. The one or morebifunctional tags can include one or more bifunctional antibodies. Theone or more bifunctional antibodies can be configured to bind to one ormore of the target component and the reactive component. The one or moretarget components can include one or more of circulating target cells orcirculating target emboli. The one or more target components caninclude, but is not limited to, one or more of tumor cells, emboli,misfolded proteins, aggregated proteins, antibodies, autoimmuneantibodies, infectious agents, or infected cells. The one or more targetcomponents can include, but is not limited to, cancer cells, pre-cancercells, autoimmune-related cells, B cells, T cells, phagocytes,platelets, lipoproteins, parasites, viruses, bacteria, fungi, orinfected cells.

The one or more reactive components can include one or more of adenaturing agent, degradative agent, or binding agent. The one or morereactive components can further include a cytotoxic agent, a cytostaticagent, a programmed cell death-inducing agent, a chemotherapeutic agent,or an antibody-toxin agent. A programmed cell death-inducing agent caninclude an agent or compound that induces programmed cell death ineukaryotic or prokaryotic cells, e.g., bacterial cells, or an agent orcompound that induces apoptosis in eukaryotic cells, e.g., parasitecells, pre-cancer cells, cancer cells, or other vertebrate cells.

A method for treating an disease, condition, or infection in avertebrate subject is described that includes providing an implantabledevice including a body defining at least one lumen configured for fluidflow; at least one first reservoir disposed within the at least onelumen and configured to include one or more bifunctional tags, whereinthe one or more bifunctional tags are configured to selectively bind toone or more target components in one or more of blood fluid or lymphfluid of a vertebrate subject; at least one treatment region disposedwithin the at least one lumen; and at least one second reservoirdisposed in the at least one treatment region and configured to includeone or more reactive components, wherein the one or more reactivecomponents are configured to sequester the one or more bifunctional tagswhen bound to the one or more target components.

Diseases, conditions, or infections wherein the disease, condition, orinfection can be modulated, alleviated, treated, prevented, reduced oreliminated by modulating a physiological effect of one or more targetcomponents in the blood fluid or lymph fluid include, but are notlimited to, cardiovascular diseases (e.g., ischemic heart disease,inflammatory heart disease), metabolic diseases (e.g., diabetes),gastrointestinal diseases (e.g., colitis, Crohn's disease), bacterialinfections (e.g., Staphylococcus bacteremia, anthrax), viral infections(e.g., AIDS, hepatitis, hemorrhagic fever), parasitic infections (e.g.,malaria, sleeping sickness, Chagas disease), metastatic cancer (e.g.,lung, breast, skin, colon, kidney, prostate, pancreas, and cervix);blood cancers (e.g., leukemia, lymphoma, Hodgkin's disease, myeloma);chemical or biological agent exposure (e.g., drug overdose, poisoning,exposure to environmental toxin). Additional examples include a numberof inflammatory diseases including but not limited to systemicinflammatory response syndrome, sepsis, septic shock, multiple organdysfunction syndrome, ischemia reperfusion, hyperreactive airwaydisease, (e.g., asthma, chronic obstructive pulmonary disease, rhinitis,sinusitis), allergic reaction, anaphylaxis, autoimmune disease,infectious disease, pulmonary failure, allograft rejection, graft versushost disease (GVHD), chronic inflammatory disease, psoriatic arthritis,rheumatoid arthritis.

A system is described that includes an implantable device including abody defining at least one lumen configured for fluid flow; at least onefirst reservoir disposed within the body and configured to include oneor more bifunctional tags, wherein the one or more bifunctional tags areconfigured to selectively bind to one or more target components in oneor more of blood fluid or lymph fluid of a vertebrate subject; at leastone treatment region disposed within the at least one lumen; and atleast one second reservoir disposed in the at least one treatment regionand configured to include one or more reactive components, wherein theone or more reactive components are configured to sequester the one ormore bifunctional tags when bound to the one or more target components.The one or more reactive components can have an increased affinity forthe one or more bifunctional tags bound to the one or more targetcomponents compared to an affinity for the one or more bifunctional tagsunbound to the one or more reactive components. The one or more reactivecomponents can be configured to modulate a physiological effect of theone or more target components. The one or more reactive components caninclude, but is not limited to, a one or more of a denaturing agent,degradative agent or binding agent.

With reference to the figures, and with reference now to FIGS. 1, 2, and3, depicted is an aspect of a device, system, or method that can serveas an illustrative environment of and/or for subject mattertechnologies, for example, device including one or more first reservoirsconfigured to include one or more bifunctional tags configured tofunction in, or proximal to, one or more blood vessel or lymph vessel ofa vertebrate subject and configured to selectively bind to one or moretarget components in one or more of blood fluid or lymph fluid of thevertebrate subject; and one or more second reservoirs configured toinclude one or more reactive components configured to sequester the oneor more bifunctional tags when bound to the one or more targetcomponents. The specific devices and methods disclosed herein areintended as merely illustrative of their more general counterparts.

Referring to FIG. 1, depicted is a partial diagrammatic view of anillustrative embodiment of an implantable device 100 including a bodydefining at least one lumen 110 configured for fluid flow; at least onecontrollable flow barrier 120 to fluid flow into the at least one lumen110; at least one first reservoir 130 disposed within the body andconfigured to include one or more bifunctional tags 140, wherein the oneor more bifunctional tags 140 are configured to selectively bind to oneor more target components 185 in one or more of blood fluid or lymphfluid 190 of a vertebrate subject; at least one treatment region 150disposed within the at least one lumen 110; and at least one secondreservoir 160 disposed in the at least one treatment region 150 andconfigured to include one or more reactive components 170, wherein theone or more reactive components 170 are configured to sequester the oneor more bifunctional tags 140 when bound to the one or more targetcomponents 185. The one or more reactive components 170 include, but arenot limited to, a denaturing agent, a degradative agent, or an energysource, wherein the one or more reactive components 170 is configured toalter, arrest, or destroy the one or more target components 85. Thedevice can further include one or more sensors 180 configured to detectone or more of unbound bifunctional tags or bifunctional tags 140 boundto target components 185 in the one or more of blood fluid or lymphfluid 190 of the vertebrate subject. The device can further include atleast one controller 195 in communication with the one or more sensor180 and in communication with the at least one controllable flow barrier120 to the at least one lumen 110, wherein the one or more reactivecomponents 170 is configured to contact the one or more of blood fluidor lymph fluid 190 of the vertebrate subject.

Referring to FIG. 2, depicted is a partial diagrammatic view of anillustrative embodiment of calculations of a target value of one or moretarget components in an implantable device including one or more sensorconfigured to detect one or more of unbound bifunctional tags orbifunctional tags bound to the target components in the one or more ofblood fluid or lymph fluid of the vertebrate subject and including atleast one controller in communication with the one or more sensor and incommunication with at least one controllable flow barrier to at leastone lumen. In an aspect, the target value can include a desiredconcentration of the one or more target components in the peripheralblood, or the target value can include a desired range of concentrationsof the one or more target components in the peripheral blood. In afurther aspect, the target value can include a desired ratio ofconcentrations of two or more target components in the peripheral blood.In an aspect, the target value can be used to determine relative levelsof the target components. The desired ratio of concentrations can bedetermined by any method or means, including for example, by a leastsquares fit of the concentrations of the two or more target components.For example, the desired ratio of concentrations can be determined by aleast squares fit of the concentrations of the two or more targetcomponents at concentrations x₁, x₂, x₃, and x₄ for a first inflammatorymediator, X, and at concentrations y₁, y₂, y₃, and y₄ for a secondinflammatory mediator, Y. The least squares can fit to a line or to atwo or three dimensional space indicating the preferred ratio of the twoor more target components.

Referring to FIG. 3, depicted is a logic flowchart of a method 301 fortreating an inflammatory disease or inflammatory condition in a subject.The method includes providing an implantable device 302 including a bodydefining at least one lumen configured for fluid flow; at least onecontrollable flow barrier to fluid flow into the at least one lumen; atleast one first reservoir disposed within the body and configured toinclude one or more bifunctional tags, wherein the one or morebifunctional tags are configured to selectively bind to one or moretarget components in one or more of blood fluid or lymph fluid of avertebrate subject; at least one treatment region disposed within the atleast one lumen; and at least one second reservoir disposed in the atleast one treatment region and configured to include one or morereactive components, wherein the one or more reactive components areconfigured to sequester the one or more bifunctional tags when bound tothe one or more target components. In an aspect, the one or morereactive components 303 can have an increased affinity for the one ormore bifunctional tags bound to the one or more target componentscompared to an affinity for the one or more bifunctional tags unbound tothe one or more reactive components. In an aspect, the one or morereactive components 304 can include a one or more of a denaturing agent,degradative agent, binding agent, or energy source.

Target Components

The device disclosed herein is configured to remove one or more targetcomponents from the blood fluid or lymph fluid of a vertebrate subjectusing one or more bifunctional tags. The target component can be anormal component or an abnormal component of the blood fluid or lymphfluid of the vertebrate subject. The target component can be associatedwith a normal physiological state or with a pathological state of thesubject. The target component can be a non-cellular component or acellular component of the blood fluid or lymph fluid of a vertebratesubject. Examples of non-cellular target components include but are notlimited to proteins, lipids, sugars, minerals, vitamins or combinationsthereof. Examples of cellular target components include but are notlimited to red blood cells, white blood cells, pathogens,pathogen-infected blood cells, cancer cells. In a further aspect, theone or more target components can be one or more of a blood clot, athrombus, an embolus, a plaque, a lipid, an aggregate, a cell, aspecific type of cells, a cell fragment, a cellular component, anorganelle, a collection or aggregation of cells, a therapeutic agent, anillicit drug, a drug of abuse, a toxin.

In an aspect, the one or more target components are cells circulating inthe blood and/or lymph of a vertebrate subject. Cellular targetcomponents can include but are not limited to blood cells (e.g.,platelets, red blood cells, neutrophils, lymphocytes, monocytes,eosinophils, basophils), pathogens (e.g., virus, bacteria, fungus,parasite), and cancer cells (e.g., metastatic cancer cells, blood cancercells).

The one or more cellular target components can be one or more bloodcells associated with a pathological state in which the normalcirculating levels of one or more class of blood cells is elevated. Forexample, elevated levels of red blood cells are associated with exposureto carbon monoxide, long-term lung disease, kidney disease, somecancers, certain forms of heart disease, liver disease. Elevated levelsof platelets are associated with bleeding, iron deficiency, cancer, orbone marrow pathologies. Elevated levels of neutrophils and eosinophilsare associated with infection, malignancy and autoimmune diseases. In anaspect, the cellular target components are blood cells that are modifiedor altered as a result of a disease, condition and/or infection. Forexample, hyperactivated B-lymphocytes in patients with inflammatorybowel disease exhibit increased surface expression of toll-like receptor2 (TLR2) relative to B-lymphocytes from normal individuals. See, e.g.,Noronha, et al., J. Leukoc. Biol. 86: Epub ahead of print; Rea, WebMD.Complete Blood Count (CBC) atwww.webmd.com/a-to-z-guides/complete-blood-count-cbc. Last updated Sep.12, 2008; accessed Oct. 5, 2009; each of which is incorporated herein byreference.

The one or more cellular target components can include one or morepathogens or pathogen-infected cells circulating in the blood and/or thelymph of a vertebrate subject. Examples of blood borne pathogens includebut are not limited to viruses, e.g., human immunodeficiency virus(HIV), and the hepatitis B, hepatitis C, and hepatitis D viruses,;bacteria, e.g., Staphylococcus, Streptococcus, Pseudomonas, Haemophilus,Escherichia coli; fungi, e.g., Aspergillus, Candida albicans, Candidaglabrata, Torulopsis glabrata, Candida tropicalis, Candida krusei, andCandida parapsilosis; and parasites, e.g., Trypanosoma cruzi,Trypanosoma brucei, Leishmania, Plasmodium, Babesia microti, Toxoplasmagondii. Other bacterial pathogens that might be found in the bloodand/or lymph at some point during a bacterial infection include but arenot limited to Bartonella, Coxiella burnetii, Chlamydia, Salmonella,Shigella, Yersinia, Legionella, Neisseria, Mycobacterium tuberculosis,Listeria, Corynebacterium diphtheriae, Campylobacter, Enterobacter.Other viral pathogens or pathogen-infected cells that might be found inthe blood and/or lymph at some point during a viral infection include,but are not limited to, cells infected with cytomegalovirus, influenza,human T-lymphotrophic virus, Epstein-Barr virus, roseolovirus, herpeslymphotropic virus, Karposi's sarcoma-associated herpesvirus, herpessimplex virus, Ebola virus, Marburg virus.

In an aspect, the one or more cellular target components can include oneor more circulating blood cells infected with a pathogen including butnot limited to bacteria, virus, or parasite. In an aspect, the one ormore cellular target components are circulating blood cells infectedwith bacteria such as, for example, red blood cells infected with B.bacilliformis or Bartonella spp. See, e.g., Dehio, Cell. Microbiol. 10:1591-1598, 2008; Chomel et al., Vet. Res. 40: 29, 2009, each of which isincorporated herein by reference. In an aspect, the one or more cellulartarget components are one or more cells infected with HIV, primarilyCD4+ T lymphocytes but also including macrophages and dendritic cells.In a further aspect, the one or more cellular target components are redblood cells infected with the malaria parasite Plasmodium falciparum.Red blood cells infected with P. falciparum can be distinguished fromnormal red blood cells by visual inspection, changes in granularitiesand changes in surface protein expression including expression on thered blood cell surface of the parasite derived protein P. falciparumerythrocyte membrane protein (PfEMP1). See, e.g., Dempster & Di Ruperto,Circuits and Systems, 2001. ISCAS 2001, The 2001

IEEE International Symposium, 5: 291-294, 2001; Weatherall, et al.,Hematology Am. Soc. Hematol. Educ. Program 35-57, 2002; Horata, et al.,Malaria J. 8:184, 2009, each of which is incorporated herein byreference.

The one or more cellular target components can include one or morecancer cells circulating in the blood and/or lymph of a vertebratesubject. In an aspect, the cancer cells can be circulating tumor cellsthat have metastasized from solid tumors located elsewhere in the body.Examples of solid tumors from which metastatic cells can arise includebut are not limited to carcinomas (e.g., adrenal, breast, cervical,colon, endometrial, lung, ovarian, pancreatic, prostate, stomach,testicular, thyroid, melanoma, head & neck) and sarcomas (e.g., brain,Ewing's sarcoma, Karposi's sarcoma, osteosarcoma, spinal cord).Circulating tumor cells are indicative of metastasis and may suggest aneed for changes in the treatment regime. For example, the detection ofcirculating tumor cells in melanoma patients who are clinically“disease-free” indicates disease recurrence, tumor cell spreading, and ahigh potential for distant metastasis, and enables identification ofhigh-risk melanoma patients. See, e.g., Schuster et al., Clin. CancerRes. 13:1171-1178, 2007, which is incorporated herein by reference. Theappearance of circulating tumor cells may also provide an indication ofthe long term prognosis for the patient. For example, breast cancerpatients with levels of circulating tumor cells equal to or higher thanfive cells per 7.5 milliliters of blood have a shorter medianprogression-free survival (2.7 months vs. 7.0 months) and shorteroverall survival (10 .1 months versus greater than 18.0 months) ascompared with breast cancer patients with less than five cells per 7.5milliliters of blood. See, Cristofanilli et al. N. Engl. J. Med.351:781-791, 2004, which is incorporated herein by reference.

In an aspect, the cancer cells can include associated with bloodcancers. Examples of blood cancers include but are not limited tolymphoma, various types of leukemia, and multiple myeloma. Lymphoma is acancer of lymphocytes which usually begins in a lymph node but canoriginate from the stomach, intestines, skin or any other organ. The twomain types of lymphoma are Hodgkin's disease and non-Hodgkin's lymphoma.In Hodgkin's disease, the abnormal cells are called the Reed-Sternbergcells, giant binucleated or multinucleated macrophages. This type ofcancer can spread throughout the lymphatic system, affecting any organor lymph tissue in the body. Non-Hodgkin's lymphoma is classified by thesize, type and distribution of cancer cells in the lymph nodes.Low-grade lymphomas include small-lymphocytic lymphoma, follicularsmall-cleaved-cell lymphoma, and follicular mixed-cell lymphoma.Intermediate-grade lymphomas include follicular large-cell lymphoma,diffuse small-cleaved-cell lymphoma., diffuse mixed lymphoma, anddiffuse large-cell lymphoma. High-grade lymphomas include immunoblasticlymphoma, lymphoblastic lymphoma, and small noncleaved (Burkitt's andnon-Burkitt's) lymphoma. Multiple myeloma is cancer of the bone marrowcaused by the uncontrolled growth of effector B cells. Effector B cellsnormally make antibodies (e.g., immunoglobulins) to fight infections. Inmultiple myeloma effector B cells multiply uncontrollably, generatingtoo much of a single type of immunoglobulin. The level of otherimmunoglobulins drops, leaving the patient vulnerable to infection. Thecancerous plasma cells collect in the bones and bone marrow and can formtumors that destroy the bone tissue, causing the bones to become fragileand prone to fracture.

In an aspect, the one or more target components include non-cellularcomponents present in the blood and/or lymph of a vertebrate subject.Non-cellular components can include but are not limited to sugars (e.g.,glucose), lipids (e.g., triacylglycerols, cholesterol, phospholipids),vitamins, minerals, non-protein hormones (e.g., estrogen, testosterone),proteins (e.g., enzymes, hormones, antibodies, blood clotting factors,lipoproteins). Additional examples of proteins found in the blood and/orlymph include but are not limited to serum proteins (e.g., subclasses ofimmunoglobulins, complement factors, C1 esterase, circulating immunecomplexes, albumin, anti-trypsin, fetoprotein, acid glycoprotein,alpha-macroglobulin, beta-microglobulin, ceruloplasmin, transferrin),acute phase proteins associated with disease (e.g., C-reactive protein,SPLA2, ferritin), coagulation or complement related proteins (e.g.,tissue-factor pathway inhibitor, soluble tissue factor, kallikrein,factor XIIa, thrombin, lupus anticoagulant, soluble CD46, soluble CD55),and markers of cellular activation (e.g., elastase, elastase/antitrypsincomplexes, lactoferrin, granzym, nucleosomes, soluble CD16, solubleCD27).

The one or more target components can include one or more inflammatorymediators. Examples of inflammatory mediators include but are notlimited to interferons (IFN) IFN-α, IFN-β, and IFN-γ; interleukins (IL)IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11,IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, IL-19, IL-20, IL-21,IL-22, IL-23, IL-24, IL-27, IL-28, IL-29, IL-30, IL-31, and IL-32; tumornecrosis factor (TNF) TNF-α and TNF-β; granulocyte colony stimulatingfactor (G-CSF); granulocyte-macrophage colony stimulating factor(GM-CSF); macrophage colony-stimulating factor (M-CSF); gelsolin,erythropoietin (EPO); and thrombopoietin (TPO). The one or moreinflammatory mediators can be any of a number of chemotactic cytokines(chemokines) including but not limited to CC chemokines CCL1 throughCCL28 exemplified by RANTES (CCL5), MCP-1 (CCL2), LARC(CCL20), MIP-1a(CCL3), and MDC(CCL22); CXC chemokines CXCL1 through CXCL17 exemplifiedby LIX (CXCL5), GCP-2 (CXCL6) and BCA-1 (CXCL13); C chemokines XCL1 andXCL2; CX3C chemokine C3CL1 (fractalkine); and chemokine like moleculesexemplified by MIF. Other inflammatory mediators include but are notlimited to anaphylatoxin fragments C3a, C4a, and C5a from the complementpathway; leukotrienes LTA4, LTB4, LTC4, LTD4, LTE4, and LTF4;prostaglandins; growth factors EGF, FGF-9, FGF-basic, growth hormone,stem cell factor (SCF), TGF-β and VEGF; soluble receptors to tumornecrosis factor receptor (sTNFr); soluble interleukin receptors sIL-1rand sIL-2r; C-reactive protein; CD11b; histamine; serotonin;apolipoprotein A1; β2-microglobulin; bradykinin; D-dimer; endothelin-1;eotaxin; factor VII; fibrinogen; GST; haptoglobin; IgA; insulin; IP-10;leptin; LIF; lymphotactin; myoglobin; OSM; SGOT; TIMP-1; tissue factor;VCAM-1; VWF; thromboxane; platelet activating factor (PAF);immunoglobulins; and endotoxins such as lipopolysaccharide (LPS); andvarious exotoxins such as superantigens, e.g., from, Staphylococcusaureus and Streptococcus pyogenes.

In an aspect, the one or more target components include exogenouschemical or biological agents that have been introduced into the bloodand/or lymph of a vertebrate subject. Examples of exogenous targetcomponents include, but are not limited to, drugs, both legal andillegal, poisons, and environmental toxins. Examples of drugs commonlyused to treat disease include but are not limited to anti-depressants,anti-psychotics, anti-virals, anti-fungals, anti-parasitics,anti-protozoal drugs, anti-inflammatory, antibiotics, analgesics,anti-hypertensives, statins, other cardiovascular drugs, antiseizuredrugs, muscle relaxants, hormones, steroids, chemotherapeutic agents.Examples of common illicit drugs or drugs of abuse include but are notlimited to cannabinoids such as hashish and marijuana; depressants suchas barbiturates, benzodiazepines (e.g., Valium, Halcion), gamma hydroxybutyrate (GHB), and methaqualone; dissociative anesthetics such asketamine and phencyclidine (PCP); hallucinogens such as LSD, mescaline,ibogaine, and psilocybin; opioids and morphine derivatives such ascodeine, fentanyl, heroin, morphine, opium, oxycodone (OxyContin) andhydrocodone bitartate/acetaminophen (Vicodin); stimulants such asamphetamines, methamphetamine, cocaine, methylphenidate (Ritalin), MDMA(ecstasy), and nicotine; and anabolic steroids (see “Commonly AbusedDrugs”, National Institute on Drug Abuse, www.drugabuse.gov). Examplesof environmental toxins include but are not limited to lead, arsenic,mercury, phthalates. Examples of additional environmental toxins can befound in ATSDR: Safeguarding Communities from Chemical Exposures,Centers for Disease Control, and in the Agency for Toxic Substances &Disease Registry as part of the Centers for Disease Control,www.atsdr.cdc.gov; Wang. J. Med. Toxicol. 4:143-4, 2006, each of whichis incorporated herein by reference.

Controlling Levels of One or More Target Components to a Target Value

A device is disclosed herein that includes one or more sensor configuredto detect one or more target components in the blood fluid or lymphfluid of a vertebrate subject and configured to control levels of theone or more target components to a target value. The target value can bea desired concentration of one or more target components in the bloodfluid or lymph fluid, or the target value can be a desired range ofconcentrations of one or more target components in the blood fluid orlymph fluid. Alternatively, the target value can be a desired ratio ofconcentrations of two or more target components in the blood fluid orlymph fluid. The desired ratio can be determined by a least squares fitof the concentrations of the two or more target components. The targetvalue of a target component can be a desired concentration and/orconcentration range and/or ratio of concentrations that is a specificvalue or range of values such as, for example, a value or range ofvalues observed in a normal subject. Alternatively, the target value ofa target component can be a desired concentration and/or concentrationrange and/or ratio of concentrations that is at least 20%, at least 40%,at least 60%, at least 80%, or at least 100% below or above the currentlevel of the target component in the blood fluid or lymph fluid of avertebrate subject.

The target value of one or more target components can include a desiredconcentration and/or concentration range below that observed in theblood fluid or lymph fluid of a vertebrate subject experiencing adisease, disorder, or infection. For example, a number of targetcomponents, e.g., inflammatory mediators, are elevated in the blood ofsubjects diagnosed with systemic immune response syndrome (SIRS) andsepsis. See, e.g., Ueda, et al., Am. J. Respir. Grit. Care Med.160:132-136, 1999; Kurt, et al., Mediators Inflamm. 2007:31397, 2007;Kellum, et al., Arch. Intern. Med. 167:1655-1663, 2007; Wang, et al.,Crit. Care 12:R106, 2008; each of which is incorporated herein byreference. As an example, the levels of TNF-α, IL-6, and IL-8 in normalsubjects is reported as less than 5 pg/ml, less than 10 pg/ml, and lessthan 10 pg/ml, respectively. In individuals with septic shock, the serumlevels of TNF-α, IL-6, and IL-8 are significantly elevated to meanvalues of 138+/−22 pg/ml, 27,255+/−7,895 pg/ml, and 2,491+/−673 pg/ml,respectively. See Ueda, et al., Am. J. Respir. Crit. Care Med.160:132-136, 1999; which is incorporated herein by reference.

The relative levels of one or more target components in the blood fluidor lymph fluid of a vertebrate subject can be correlated with prognosisand survival. For example, sepsis non-survivors have proportionallyhigher levels of inflammatory mediators relative to sepsis survivors andnormal controls. In one study, high levels of both IL-10 (mean of 45pg/ml) and IL-6 (mean of 735 pg/ml) at hospital admission wereassociated with increased mortality as compared with low initial levelsof IL-10 (mean of 7.4 pg/ml) and IL-6 (mean of 15 pg/ml). In anotherexample, elevated serum levels of IL-6 were correlated with sepsissymptom scores and poor outcome. These data suggest that modulating thelevels of one or more inflammatory mediators to a desired target valuein the blood can alter the course of the disease. See, e.g., Kellum, etal., Arch. Intern. Med. 167:1655-1663, 2007; Presterl, et al., Am. J.Respir. Crit. Care Med. 156:825-832, 1997; each of which is incorporatedherein by reference.

The relative levels of one or more target components in the blood fluidor lymph fluid of a vertebrate subject can be correlated with a chronicdisease. For example, subjects with rheumatoid arthritis have increasedlevels of various inflammatory mediators relative to normal subjectsincluding IL-6 (15.8 pg/ml versus 4.0 pg/ml), TNF-α (10 pg/ml versus 3.2pg/ml), IL-1β (129.8 pg/ml versus 57.3 pg/ml), IL-8 (9.3 pg/ml versus2.6 pg/ml), IL-10 (15.5 pg/ml versus 4.6 pg/ml), and IL-12 (20.2 pg/mlversus 6.2 pg/ml). Psoriatic arthritis is also characterized byincreased levels of circulating inflammatory mediators withstatistically significant increases in the serum levels of variousinflammatory mediators in subjects diagnosed with psoriatic arthritisversus normal. See, e.g., Nowlan, et al., Rheumatology 45:31-37, 2006;Mittal & Joshi, .J. Indian Rheumatol. Assoc. 10:59-60, 2002; Szodoray,et al., Rheumatology 46:417-425, 2007, each of which is incorporatedherein by reference.

The target value of one or more target components that are cells can bea desired concentration or concentration range that is below thatobserved in the blood fluid or lymph fluid of a vertebrate subjectexperiencing a disease, condition or infection. For example, elevatedlevels of red blood cells are associated with exposure to carbonmonoxide, long-term lung disease, kidney disease, some cancers, certainforms of heart disease, liver disease. Elevated levels of platelets areassociated with bleeding, iron deficiency, some diseases like cancer, orbone marrow problems. Elevated levels of neutrophils, eosinophils,and/or lymphocytes are associated with infection, malignancy andautoimmune diseases. The desired concentration or concentration rangecan be the concentration or concentration range observed in a normalindividual. For example, the normal range of white blood cells in menand nonpregnant women ranges from 4.5 to 11×10⁹ cells per liter while inpregnant women, the white blood cell counts range from 5.9 to 25.7×10⁹cells per liter depending upon whether the subject is in the first,second or third trimester or postpartum. Similarly, normal red bloodcell counts range from 4.7 to 6.1×10¹² cells per liter in men, 4.2 to5.4×10¹² cells per liter in women, 4.0 to 5.5×10¹² cells per liter inchildren and 4.8 to 7.1×10¹² cells per liter in newborns. Normalplatelet counts range from 150 to 450×10⁹ cells per liter for childrenand 150 to 400×10⁹ cells per liter for adults. See, e.g., Rea, WebMD.Complete Blood Count (CBC) atwww.webmd.com/a-to-z-guides/complete-blood-count-cbc. Last updated Sep.12, 2008; accessed Oct. 5, 2009; incorporated herein by reference.

The target value can be a percentage range of cells in the blood. Forexample, of the total white blood cells in a normal subject, neutrophilsrange from 50% to 62%, band neutrophils range from 3% to 6%, lymphocytesrange from 25% to 40%, monocytes range from 3% to 7%, eosinophils rangefrom 0% to 3%, and basophils range from 0% to 1%.

The target value can be a desired ratio of concentrations of two or moretarget components in the blood fluid or lymph fluid as determined by aleast squares fit of the concentration values of the two or more targetcomponents. In this instance, the levels of one or more targetcomponents can be altered to modulate the overall ratio of two or moretarget components. For example, levels of neutrophils relative toleukocytes is reportedly is correlated with cardiovascular risk in thatincreased neutrophils and/or decreased leukocytes are associated withdiabetes, coronary artery disease, unstable angina, and increased riskof myocardial infarction. See, e.g., Home, et al., J. Am. Coll. Cardiol.45: 1638-1643, 2005, which is incorporated herein by reference.

In some pathological states such as cancer or infection, the idealtarget value of one or more target components can be zero. In theinstance where a target value of zero is not attainable, the targetvalue can be a value that reduces the symptoms and/or the diseaseprogression. In malaria infected individuals, for example, the degree ofparasitemia is correlated with the severity of the disease. The numberof parasites per microliter of blood is used to assess parasitemia. Forexample, a subject may just be showing signs of symptoms at 100parasites per microliter (0.002% parasitemia), severe malaria at 100,000to 250,000 parasites per microliter (2-5% parasitemia), and near deathat 500,000 parasites per microliter (10% parasitemia). Reducingparasitemia can reduce symptoms and disease severity.

Similarly, the target value of one or more target component that is atoxin or illicit drug can be zero. In the instance where a target valueof zero is not attainable, the target value can be a value that reducestoxicity. For example, elevated levels of lead in the blood in adultscan damage the nervous, hematologic, reproductive, renal,cardiovascular, and gastrointestinal systems. The majority of cases oflead poisoning are workplace related. The U.S. Department of Health andHuman Services recommends that blood levels of lead among adults bereduced to <25 ug/dL. The highest blood levels of lead acceptable bystandards of the U.S. Occupational Safety Health Administration is 40ug/dL. The geometric mean blood levels of lead of all adults in the USis <3 ug/dL. MMWR 58(14):365-369, 2009, which is incorporated herein byreference.

Device Functioning in or Proximal to Blood and/or Lymph Vessel of aVertebrate Subject

A device is disclosed herein for treating a disease, condition orinfection including at least one first reservoir that includes one ormore bifunctional tags to selectively bind to one or more targetcomponents in one or more of blood fluid or lymph fluid of a vertebratesubject and at least one second reservoir disposed in at least onetreatment region and configured to include one or more reactivecomponents, wherein the one or more reactive components are configuredto sequester the one or more bifunctional tags when bound to the one ormore target components. The device can include one or more sensorconfigured to sense one or more target components in the blood fluid orlymph fluid of the vertebrate subject, a controller in communicationwith and responsive to the sensor, and a means for releasing one or morebifunctional tags from a first set of reservoirs responsive to thecontroller. The one or more bifunctional tags can be released into theblood fluid or lymph fluid of a vertebrate subject and are configured tobind one or more target components to form a bifunctional tag/targetcomponent complex. The controller can controllably adjust the release ofthe bifunctional tags to achieve a target value of the detected one ormore target components in the blood of the subject. The device canfurther include a second set of reservoirs configured to bind,sequester, and optionally inactivate the bifunctional tag/targetcomponent complex. The means for sequestering and optionallyinactivating the bifunctional tag/target component complex can includeone or more second reservoirs configured to receive at least a portionof the blood fluid or lymph fluid through a flow route, the controllerconfigured to control flow of blood fluid or lymph fluid through theflow route into the second reservoir, and the second reservoir includingone or more reactive components configured to sequester and optionallyinactivate the bifunctional tag/target component complex. The controllercan control at least a portion of the blood fluid or lymph fluid througha flow route in communication with and responsive to a tag sensorconfigured to detect the bifunctional tag/target component complex.

The device for binding and sequestering one or more target componentsassociated with a disease, condition or infection is in whole, or inpart, configured for use in, or proximal to, one or more blood vesselsand/or lymph vessels of a vertebrate subject. In an aspect, the devicein part or in whole is an intra-vessel sized device (e.g., sufficientlysmall enough to be placed in a blood vessel and/or a lymph vessel whilenot necessarily obstructing flow). The device can be inserted into ablood vessel or lymph vessel. Configurations for the device include, butare not limited to, a substantially tubular structure, with one or morelumens in fluid communication with the blood vessel or lymph vessel of avertebrate subject. In a further aspect, the device can take the form ofa short cylinder, an annulus, a cylinder, and/or a spiral. See, e.g.,U.S. Patent Applications 2007/0066929 and 2008/0058785; Bezrouk et al,Scripta Medica (BRNO) 78(4):219-226, 2005, each of which is incorporatedherein by reference. In an aspect, the device has a cylindrical andhollow configuration, with a single central opening, optionally allowingthe exterior of the cylindrical structure to contact and engage the wallof the vessel, and the interior of the structure (within the singlecentral opening) to form a fluid-contacting portion of the device. Forexample, the device can be configured as a specialized stent fixedwithin a specific artery or vein. See, e.g., U.S. Pat. Nos. 5,411,551,7,326,240; U.S. Patent Applications 2007/0294150, 2008/0281400; Yokota,et al., 22nd IEEE International Conference MicroElectro MechanicalSystems, Sorrento, Italy, January 25-29. IEEE pp. 495-499, 2009, each ofwhich is incorporated herein by reference.

In an aspect, the device in whole or in part is configured to beapproximately hemi-spherical or hemi-elliptoid, allowing a portion ofits cross-section to contact and/or engage the internal wall of a bloodvessel or lymph vessel without significantly and/or substantiallyobstructing the movement of fluid within the vessel. The device caninclude one or more wall engaging components including, but are notlimited to, rotating wheels, projections (e.g. arms), springs, hooks(e.g. claws), suction cups, and/or tissue adhesives that are configuredto engage wall portions.

In an aspect, the device can be configured in a pill- or capsule-shape,and configured to move through a central portion of a vessel. The devicecan engage a wall of the vessel using one or more engaging componentsand/or freely travel through the blood and/or lymph systems. See, e.g.,U.S. Patent Application 2007/0156211 A1, which is incorporated herein byreference. The device can be targeted to a site of disease (e.g.,inflammation, cancer) in the subject. In an aspect, the device can senseelevated levels of one or more target components in the blood orlymphatic system of the subject and can target and form a stationarylocation at, or near, a site of disease, condition or infection in theperipheral circulation of the subject. In an aspect, the implantabledevice can be incorporated into a shunt, for example, an arteriovenousshunt inserted between an artery and a vein.

In an aspect, the device in part or in whole is positioned proximal to ablood vessel or lymph vessel. “Proximal to” can refer to a space or areanear to a blood vessel or lymph vessel. Locations that are proximal to avessel can include, for example, locations external to the vessel wallwhere there is space for implanting one or more devices in whole or inpart, and optionally to facilitate external access to the devices inwhole or in part. In an aspect, “proximal to” can include distances suchas, but not limited to, approximately 0.1, 1.0, 10, and/or 100 μmsand/or approximately 0.1, 1.0, 10, and/or 100 mms, and can optionallyinclude larger and/or smaller distances depending on, for example, theavailability of space and the size of the device and/or the vessel.

In an aspect, the device is configured as a self-contained unit thatincludes all functionalities necessary for operation of the device. Inan aspect, the system can be configured as one or more components in oneor more locations separate from one another, wherein one or more of thecomponents includes one or more essential and/or non-essentialfunctionalities. As an example, one component of the system can beplaced within a blood vessel, and another component of the system placedproximal to the blood vessel optionally in a location more accessiblefrom the exterior of the subject, or where there is additional space. Aremote portion can be configured to provide for monitoring of the vesselportion of the system, data collection, or data analysis, and/orremote-control of one or more other functions of the system such assensing target components, controlling flow through a flow route, andreleasing a reactive component. The remote portion can be at a separatelocation within the body of the subject, or outside the body of thesubject. Data and/or power signals can be transmitted between the one ormore components of the device using electromagnetic signals, orelectrical or optical links.

The dimensions and mechanical properties (e.g., rigidity) of the devicein part or in whole are configured for compatibility with the locationof use in order to provide for reliable positioning and/or to providefor movement of the device while preventing damage to the vessel, thevessel lumen, and/or internal location and its surrounding structure.The choice of structural component size and configuration appropriatefor a particular blood vessel or lymph vessel location can be selectedby a person of skill in the art, optionally a medical professional.Structural components of the device can be constructed using a varietyof manufacturing methods, from a variety of biocompatible materials.Appropriate materials include metals, ceramics, polymers, and compositematerials having suitable biocompatibility, sterilizability, mechanical,and physical properties. Examples of materials and selection criteriaare described, for example, in The Biomedical Engineering Handbook(Second Edition, Volume I, J. D. Bronzino, Ed., Copyright 2000, CRCPress LLC, pp. IV-1-43-22), which is incorporated herein by reference.Manufacturing techniques can include injection molding, extrusion,die-cutting, rapid-prototyping, etc., and will depend on the choice ofmaterial and device size and configuration. Sensing and energy-emittingportions of the devices as well as associated control circuitry can befabricated on the structural elements using various microfabricationand/or MEMS techniques or can be constructed separately and subsequentlyassembled to the structural elements, as one or more distinctcomponents. See, e.g., U.S. Patent Applications 2005/0221529,2005/0121411, 2005/0126916, 2007/0066939, 2007/0225633 and Nyitrai, etal. “Preparing Stents with Masking & Etching Technology” 26^(th)International Spring Seminar on Electronics Technology pp. 321-324,IEEE, 2003, each of which is incorporated herein by reference.

In additional to biocompatible materials described and incorporatedherein, flexible material having adjustable diameter, taper, and lengthproperties can be used as part of the structural material. For example,some materials can change from a longer, narrower configuration, to ashorter, wider configuration, or can taper over their length, e.g.,shape-memory polymers that can move from one shape to another inresponse to a stimulus such as heat. Structural elements that canexhibit this type of expansion/contraction property can includeself-expanding material, resilient material, and/or mesh structuresformed of various metals, e.g., ionic polymer-metal composites (IPMC) orplastics, and some polymeric materials, e.g., hydrogels, nitinol, orpolyester. See, e.g. Bellin et al., Proc. Natl. Acad. Sci. USA.103:18043-18047, 2006; and Shahinpoor & Kim, Smart Mater. Struct.10:819-833, 2001, each of which are incorporated herein by reference.

Sensing Target Components

The device includes one or more sensors for qualitatively and/orquantitatively measuring one or more target components in the bloodfluid or lymph fluid of a vertebrate subject. The one or more sensorscan include, but are not limited to, one or more of a biosensor, achemical sensor, a physical sensor, an optical sensor, or a combinationthereof. The one or more sensors can include one or more targetrecognition elements that recognize one or more target components. Theinteraction of one or more target components with one or more targetsensors results in one or more detectable signals sent to thecontroller. In response to the detectable signal, the controllercontrols the release of one or more bifunctional tags from the one ormore first reservoirs. Preferably the one or more target sensors measurein real-time the relative number of one or more target components in theblood fluid or lymph fluid of a vertebrate subject

The one or more sensors are configured to sense or detect one or moretarget components in the blood fluid or lymph fluid of a vertebratesubject. In an aspect, the one or more sensors are configured to senseone or more target components that are one or more cells in the bloodfluid or lymph fluid of a vertebrate subject. A target componentincludes a cell, for example, a bacterium, a protozoan, a platelet, ared blood cell, a lymphocyte, a monocyte, a neutrophil, an eosinophil, acirculating tumor cell, or a combination thereof. The one or more targetcomponents can be sensed using any of a number of imaging or opticalmethods including but not limited to light scattering, electricalimpedance, infrared spectroscopy, acoustic imaging, thermal imaging,photothermal imaging, dark field, visible light absorption andrefraction, and autofluorescence. See, e.g., U.S. Patent Application2009/0093728; Doornbos et al. Cytometry 14:589-594, 1993; Gao et al.Proceedings of the 25^(th) Annual International Conference of the IEEEEMBS, Cancun, Mexico, Sep. 17-21, 2003; Oberreuter et al. Int. J. Syst.Evol. Microbiol. 52:91-100, 2002; Baddour et al. Ultrasonics SymposiumIEEE 2:1639-1644, 2002; Zharov et al. J. Cell. Biochem. 97:916-932,2006; Zharov et al. J. Biomed. Opt. 11:054034-1-4, 2006; Koenig et al.J. Fluoresc. 4:17-40, 1994; which are each incorporated herein byreference. As an example, red blood cells infected with the parasitePlasmodium falciparum can be differentiated from other cells in theblood using differential light scatter at 10 degrees (complexity) andpolarized light scatter at 90 degrees (lobularity) based on thepigmentation of the parasite. See, e.g., Mendelow et al. Br. J.Haematology 104: 499-503, 1999, which is incorporated herein byreference.

In some instances, one or more target components in the blood fluid orlymph fluid of a vertebrate subject are recognized based on a spectralanalysis. Alternatively, the one or more target components arerecognized based on pattern and image recognition analysis. Variousmethods have been described for image and shape analysis of cells andsubcellular components of cells. See, e.g., U.S. Pat. Nos. 5,107,422;5,790,691; 6,956,961 B2; 7,151,847 B2; U.S. Patent Applications2005/0251347 A1; 2006/0039593 A1; Fei-Fei et al. IEEE Transactions onPattern Analysis and Machine Intelligence 28:594-611, 2006; Martin etal. IEEE Transactions on Pattern Analysis and Machine Intelligence26:530-549, 2004; Olson et al. Proc. Natl. Acad. Sci. USA 77:1516-1520,1980; Schneider, et al Biorheology 32:237-238, 1995; each of which areincorporated herein by reference. For example, a Texture AnalyzingSystem can be used to distinguish various cells in the blood and/orlymph of a vertebrate subject based on the granularity of the cell orcells. See, e.g., Bins et al. Cytometry 1:321-324, 1981, which isincorporated herein by reference. The imaged components of the cells aremeasured with a gray scale with 33 intervals ranging from black (level0) to white (level 99) and a histogram is generated. Mature white bloodcells (neutrophils, eosinophils, basophils and lymphocytes) have a densenuclear structure and therefore low counts. In contrast, monocytes havea looser, less dense nuclear structure and high counts. The cytoplasm ofeosinophils and neutrophils is very granular and is reflected in thecombination of high positive and low negative counts. Smaller values areseen in the cytoplasm of lymphocytes, monocytes and basophils.Similarly, granulometries can be used to identify red blood cellsinfected with the malarial parasite. See, e.g., Dempster & DiRubertoCircuits and Systems, 2001. ISCAS 2001. The 2001 IEEE InternationalSymposium on May 6-9, 2001, 5:291-294, which is incorporated herein byreference.

In an aspect, the one or more sensors can include one or more targetrecognition elements that recognize one or more target components. Thetarget recognition elements are configured to specifically bind one ormore target components. The target recognition elements can include, butare not limited to, antibodies, antibody fragments, peptides,oligonucleotides, DNA, RNA, aptamers, protein nucleic acids, proteins,viruses, enzymes, receptors, bacteria, cells, cell fragments, inorganicmolecules, organic molecules, or combinations thereof. The one or moretarget recognition elements can be associated with one or more substrateintegrated into the one or more target sensors. Binding of a targetcomponent to a specific target recognition element activates the sensor.

In an aspect, the one or more target sensors can use Förster orfluorescence resonance energy transfer (FRET) to sense one or moretarget components in the blood fluid or lymph fluid of a vertebratesubject. FRET is a distance-dependent interaction between the electronicexcited states of two dye molecules in which excitation is transferredfrom a donor molecule to an acceptor molecule without emission of aphoton. In an aspect, interaction of a donor molecule with an acceptormolecule results in a shift in the emission wavelength associated withexcitation of the acceptor molecule. In other aspects, interaction of adonor molecule with an acceptor molecule in results in quenching of thedonor emission. The one or more target recognition elements associatedwith the one or more sensors can include at least one donor molecule andat least one acceptor molecule. Binding of a target component to thetarget recognition element results in a conformation change in thetarget recognition element, leading to changes in the distance betweenthe donor and acceptor molecules and changes in measurable fluorescence.The target recognition element can be a cell, an antibody, an aptamer, areceptor or any other molecule that changes conformation or signaling inresponse to binding a target.

A variety of donor and acceptor fluorophore pairs can be considered forFRET associated with the target recognition element including, but notlimited to, fluorescein and tetramethylrhodamine; IAEDANS andfluorescein; fluorescein and fluorescein; and BODIPY FL and BODIPY FL. Anumber of Alexa Fluor (AF) fluorophores (Molecular Probes-Invitrogen,Carlsbad, Calif., USA) can be paired with other AF fluorophores for usein FRET. Some examples include, but are not limited, to AF 350 with AF488; AF 488 with AF 546, AF 555, AF 568, or AF 647; AF 546 with AF 568,AF 594, or AF 647; AF 555 with AF594 or AF647; AF 568 with AF6456; andAF594 with AF 647.

The cyanine dyes Cy3, Cy5, Cy5.5 and Cy7, which emit in the red and farred wavelength range (>550 nm), offer a number of advantages forFRET-based detection systems. Their emission range is such thatbackground fluorescence is often reduced and relatively large distances(>100 Å) can be measured as a result of the high extinction coefficientsand good quantum yields. For example, Cy3, which emits maximally at 570nm and Cy5, which emits at 670 nm, can be used as a donor-acceptor pair.When the Cy3 and Cy5 are not proximal to one another, excitation at 540nm results in the emission of light by Cy3 at 590 nm only. In contrast,when Cy3 and Cy5 are brought into proximity by a conformation change inan aptamer, antibody, or receptor, excitation at 540 nm results in anemission at 680 nm. Semiconductor quantum dots (QDs) with variousexcitation/emission wavelength properties can also be used to generate afluorescence based sensor.

Quenching dyes are used as part of the binder element to quench thefluorescence of visible light-excited fluorophores. Examples include,but are not limited, to DABCYL, the non-fluorescing diarylrhodaminederivative dyes QSY 7, QSY 9 and QSY 21 (Molecular Probes, Carlsbad,Calif., USA), the non-fluorescing Black Hole Quenchers BHQ0, BHQ1, BHQ2,and BHQ3 (Biosearch Technologies, Inc., Novato, Calif., USA) and Eclipse(Applera Corp., Norwalk, Conn., USA). A variety of donor fluorophore andquencher pairs can be considered for FRET associated with the targetrecognition element including, but not limited to, fluorescein withDABCYL; EDANS with DABCYL; or fluorescein with QSY 7 and QSY 9. Ingeneral, QSY 7 and QSY 9 dyes efficiently quench the fluorescenceemission of donor dyes including blue-fluorescent coumarins, green- ororange-fluorescent dyes, and conjugates of the Texas Red and Alexa Fluor594 dyes. QSY 21 dye efficiently quenches all red-fluorescent dyes. Anumber of the Alexa Fluor (AF) fluorophores (MolecularProbes-Invitrogen, Carlsbad, Calif., USA) can be paired with quenchingmolecules as follows: AF 350 with QSY 35 or DABCYL; AF 488 with QSY 35,DABCYL, QSY7 or QSY9; AF 546 with QSY 35, DABCYL, QSY7 or QSY9; AF 555with QSY7 or QSY9; AF 568 with QSY7, QSY9 or QSY21; AF 594 with QSY21;and AF 647 with QSY 21.

The one or more sensor for sensing target components in the blood fluidor lymph fluid of a vertebrate subject can use the technique of surfaceplasmon resonance (for planar surfaces) or localized surface plasmonresonance (for nanoparticles). Surface plasmon resonance involvesdetecting changes in the refractive index on a sensor surface inresponse to changes in molecules bound on the sensor surface. In anaspect, the surface of the sensor is a glass support or other solidsupport coated with a thin film of metal, for example, gold. In afurther aspect, the sensor surface includes a matrix to which isimmobilized one or more target recognition elements that recognize oneor more target components. The target recognition elements can beantibodies or fragments thereof, oligonucleotide or peptide basedaptamers, receptors or ligands, artificial binding substrates formed bymolecular imprinting, or any other examples of molecules and/orsubstrates that bind cells. As blood or blood components from thesubject passes by the sensor surface, a target component can interactwith a target recognition element on the sensor surface. The sensor isilluminated by monochromatic light. Resonance occurs at a specific angleof incident light. The resonance angle depends on the refractive indexin the vicinity of the surface, which is dependent upon theconcentration of target components on the surface. An example ofinstrumentation that uses surface plasmon resonance is the BIACOREsystem (Biacore, Inc.—GE Healthcare, Piscataway, N.J.) which includes asensor microchip, a laser light source emitting polarized light, anautomated fluid handling system, and a diode array position sensitivedetector. See, e.g., Raghavan & Bjorkman Structure 3:331-333, 1995,which is incorporated herein by reference.

The one or more sensors for sensing target components can be one or morelabel-free optical biosensors that incorporate other opticalmethodologies, e.g., interferometers, waveguides, fiber gratings, ringresonators, and photonic crystals. See, e.g., Fan, et al., Anal. Chim.Acta 620:8-26, 2008, which is incorporated herein by reference.

The one or more sensors for sensing target components can be one or moremicrocantilevers. A microcantilever can act as a biological sensor bydetecting changes in cantilever bending or vibrational frequency inresponse to binding of one or more target components to the surface ofthe sensor. In an aspect the sensor can be bound to a microcantilever ora microbead as in an immunoaffinity binding array. In an aspect, abiochip can be formed that uses microcantilever bi-material formed fromgold and silicon, as sensing elements. See, e.g. Vashist J. NanotechOnline 3:DO: 10.2240/azojono0115, 2007, which is incorporated herein byreference. The gold component of the microcantilever can be coated withone or more target recognition elements which upon binding one or moretarget components causes the microcantilever to deflect. Aptamers orantibodies specific for one or more target components can be used tocoat microcantilevers. See, e.g., U.S. Pat. No. 7,097,662, which isincorporated herein by reference. The one or more sensor can incorporateone or more methods for microcantilever deflection detection including,but not limited to, piezoresistive deflection, optical deflection,capacitive deflection, interferometry deflection, optical diffractiongrating deflection, and charge coupled device. In an aspect, the one ormore microcantilever can be a nanocantilever with nanoscale components.The one or more microcantilevers and/or nanocantilevers can be arrangedinto arrays. Both microcantilevers and nanocantilevers can find utilityin microelectomechnical systems (MEMS) and/or nanoelectomechnicalsystems (NEMS).

The one or more sensors for sensing target components can be a fieldeffect transistor (FET) based biosensor. In this aspect, a change inelectrical signal is used to detect interaction of one or more targetcomponents with one or more target recognition elements of the sensor.See, e.g., U.S. Pat. No. 7,303,875, which is incorporated herein byreference.

The one or more sensors for sensing one or more target components canincorporate electrochemical impedance spectroscopy. Electrochemicalimpedance spectroscopy can be used to measure impedance across a naturaland/or artificial lipid bilayer. The sensor can incorporate anartificial bilayer that is tethered to the surface of a solid electrode.One or more receptor can be embedded into the lipid bilayer. The one ormore receptors can be ion channels that open and close in response tobinding of a specific analyte. The open and closed states can bequantitatively measured as changes in impedance across the lipidbilayer. See, e.g., Yang, et al., IEEE SENSORS 2006, EXCO, Daegu,Korea/Oct. 22-25, 2006, which is incorporated herein by reference.

The one or more sensors can include cells with binding elements whichwhen bound to target components induce a measurable or detectable changein the cells. The cells can emit a fluorescent signal in response tointeracting with one or target components. For example, a bioluminescentbioreporter integrated circuit can be used in which binding of a ligandto a cell induces expression of reporter polypeptide linked to aluminescent response. See e.g., U.S. Pat. No. 6,673,596; Durick &Negulescu Biosens. Bioelectron. 16:587-592, 2001, each of which isincorporated herein by reference. Alternatively or additionally, the oneor more cell can emit an electrical signal in response to interactingwith one or more target components. In a further aspect, an implantablebiosensor can be used which is composed of genetically modified cellsthat respond to target binding by emitting a measurable electricalsignal. See e.g., U.S. Patent Application 2006/0234369 A1; which isincorporated herein by reference.

The one or more sensors including the target recognition elements candetect one or more target components that are non-cellular targetcomponents including but are not limited to sugars (e.g., glucose),lipids (e.g., triacylglycerols, cholesterol, phospholipids), vitamins,minerals, non-protein hormones (e.g., estrogen, testosterone), proteins(e.g., enzymes, hormones, antibodies, blood clotting factors,lipoproteins). Additional examples of proteins found in the blood and/orlymph include but are not limited to serum proteins (e.g., subclasses ofimmunoglobulins, complement factors, Cl esterase, circulating immunecomplexes, albumin, anti-trypsin, fetoprotein, acid glycoprotein,alpha-macroglobulin, beta-microglobulin, ceruloplasmin, transferrin),acute phase proteins associated with disease (e.g., C-reactive protein,SPLA2, ferritin), coagulation or complement related proteins (e.g.,tissue-factor pathway inhibitor, soluble tissue factor, kallikrein,factor XIIa, thrombin, lupus anticoagulant, soluble CD46, soluble CD55),and markers of cellular activation (e.g., elastase, elastase/antitrypsincomplexes, lactoferrin, granzym, nucleosomes, soluble CD16, solubleCD27).

In an aspect, the one or more target sensors including the targetrecognition elements can detect one or more target components that areinflammatory mediators, examples of which include but are not limited tointerferons (IFN) IFN-α, IFN-β, and IFN-γ; interleukins (IL) IL-1, IL-2,IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13,IL-14, IL-15, IL-16, IL-17, IL-18, IL-19, IL-20, IL-21, IL-22, IL-23,IL-24, IL-27, IL-28, IL-29, IL-30, IL-31, and IL-32; tumor necrosisfactor (TNF) TNF-α and TNF-β; granulocyte colony stimulating factor(G-CSF); granulocyte-macrophage colony stimulating factor (GM-CSF);macrophage colony-stimulating factor (M-CSF); gelsolin, erythropoietin(EPO); and thrombopoietin (TPO). The one or more inflammatory mediatorscan be any of a number of chemotactic cytokines (chemokines) includingbut not limited to CC chemokines CCL1 through CCL28 exemplified byRANTES (CCL5), MCP-1 (CCL2), LARC(CCL20), MIP-1a (CCL3), and MDC(CCL22);CXC chemokines CXCL1 through CXCL17 exemplified by LIX (CXCL5), GCP-2(CXCL6) and BCA-1 (CXCL13); C chemokines XCL1 and XCL2; CX3C chemokineC3CL1 (fractalkine); and chemokine like molecules exemplified by MIF.Other inflammatory mediators include but are not limited toanaphylatoxin fragments C3a, C4a, and C5a from the complement pathway;leukotrienes LTA4, LTB4, LTC4, LTD4, LTE4, and LTF4; prostaglandins;growth factors EGF, FGF-9, FGF-basic, growth hormone, stem cell factor(SCF), TGF-β and VEGF; soluble receptors to tumor necrosis factorreceptor (sTNFr); soluble interleukin receptors sIL-1r and sIL-2r;C-reactive protein; CD11b; histamine; serotonin; apolipoprotein A1;β2-microglobulin; bradykinin; D-dimer; endothelin-1; eotaxin; factorVII; fibrinogen; GST; haptoglobin; IgA; insulin; IP-10; leptin; LIF;lymphotactin; myoglobin; OSM; SGOT; TIMP-1; tissue factor; VCAM-1; VWF;thromboxane; platelet activating factor (PAF); immunoglobulins; andendotoxins such as lipopolysaccharide (LPS); and various exotoxins suchas superantigens, e.g., from, Staphylococcus aureus and Streptococcuspyogenes.

In an aspect, the one or more target sensors including the targetrecognition elements can detect one or more target components that areexogenous chemical or biological agents that have been introduced intothe blood and/or lymph of a vertebrate subject. Examples of exogenoustarget components include drugs, both legal and illegal, poisons, andenvironmental toxins. Examples of drugs can include but are not limitedto anti-depressants, anti-psychotics, anti-virals, anti-fungals,anti-parasitics, anti-protozoal drugs, anti-inflammatory, antibiotics,analgesics, anti-hypertensives, statins, other cardiovascular drugs,antiseizure drugs, muscle relaxants, hormones, steroids,chemotherapeutic agents. Examples of common illicit drugs or drugs ofabuse can include but are not limited to cannabinoids such as hashishand marijuana; depressants such as barbiturates, benzodiazepines (e.g.,Valium, Halcion), gamma hydroxy butyrate (GHB), and methaqualone;dissociative anasthetics such as ketamine and phencyclidine (PCP);hallucinogens such as LSD, mescaline, ibogaine, and psilocybin; opioidsand morphine derivatives such as codeine, fentanyl, heroin, morphine,opium, oxycodone (OxyContin) and hydrocodone bitartate/acetaminophen(Vicodin); stimulants such as amphetamines, methamphetamine, cocaine,methylphenidate (Ritalin), MDMA (ecstasy), and nicotine; and anabolicsteroids (see “Commonly Abused Drugs”, National Institute on Drug Abuse,www.drugabuse.gov). Examples of environmental toxins can include but arenot limited to lead, arsenic, mercury, phthalates. Examples ofadditional environmental toxins can be found in the Agency for ToxicSubstances & Disease Registry as part of the Centers for DiseaseControl, www.atsdr.cdc.gov; Patel, J. Med. Toxicol. 4:143-4, 2006, whichis incorporated herein by reference.

In an aspect, the one or more sensors including the target recognitionelements can sense or detect one or more target components that aretarget components including, but not limited to, blood cells (e.g., redblood cells, platelets, lymphocytes, monocytes, neutrophils,eosinophils, basophils), virus-infected cells (e.g., cells infected withhuman immunodeficiency virus (HIV), hepatitis B, hepatitis C, andhepatitis D), bacteria (e.g., Staphylococcus, Streptococcus,Pseudomonas, Haemophilus, Listeria, Esherichia coli), fungi, (e.g.,Candida albicans, Candida glabrata, Aspergillus, T. glabrata, Candidatropicalis, C. krusei, and C. parapsilosis) parasites (e.g., Trypanosomacruzi, Trypanosoma brucei, Leishmania, Plasmodium, Babesia microti,Toxoplasma gondii) and cancer cells (e.g., metastatic tumor cells,hematopoietic cancer cells).

The one or more target recognition elements are configured to recognizeone or more biomolecules on the surface of the one or more targetcomponents. In an aspect, the one or more target recognition elementsare configured to recognize one or more receptor types on the surface oftarget components. Examples of receptors include but are not limited toacetylcholine receptors, adenosine receptors, adrenoceptors, GABAreceptors, angiotensin receptors, cannabinoid receptors, cholecystokininreceptors, dopamine receptors, glucagon receptors, glucocorticoidreceptors, glutamate receptors, histamine receptors, mineralocorticoidreceptors, olfactory receptors, opioid receptors, purinergic receptors,secretin receptors, serotonin receptors, somatostatin receptors, steroidhormone receptors, calcium-sensing receptor, hormone receptors,erythropoietin receptor, and natriuretic peptide receptors. Otherexamples include type I cytokine receptors (e.g., type 1 interleukinreceptors, erythropoietin receptor, GM-CSF receptor, G-CSF receptor,growth hormone receptor, oncostatin M receptor, leukemia inhibitoryfactor receptor); type II cytokine receptors (e.g., type II interleukinreceptors, interferon-α/β receptors, interferon-γ receptor); members ofthe immunoglobulin superfamily (e.g., interleukin-1 receptor, CSF1,c-kit receptor, interleukin-18 receptor); tumor necrosis factor (TNF)receptor family (e.g., TNF receptor 1 (TNF-R1), TNF receptor 2 (TNF-R2),CD27, CD40, and lymphotoxin β receptor); chemokine receptors includingserpentine CCR and CXCR receptors (e.g., CCR1 and CXCR4, andinterleukin-8 receptor); TGF β receptors. See Ozaki and Leonard, J.Biol. Chem. 277:29355-29358, 2002, which is incorporated herein byreference.

In a further aspect, the one or more target recognition elements areconfigured to recognize other biomolecules on the surface of targetcomponents including but not limited to various CD (cluster ofdesignation/cluster of differentiation) markers, intergrins, ionchannels, ATPases, cell adhesion molecules, integral membraneglycoproteins, immunoglobulins, transporters. The one or more targetrecognition elements are configured to recognize components of cellsurface biomolecules including amino acid sequence and oligosaccharidemodifications.

In an aspect, the one or more target sensors including the targetrecognition element can be configured to recognize a biomoleculeassociated with a tumor cell. Examples of tumor-associated tumor cellrecognition components can include, but are not limited to, BLySreceptor, carcinoembryonic antigen (CA-125), CD25, CD34, CD33 and CD123(acute myeloid leukemia), CD20 (chronic lymphocytic leukemia), CD19 andCD22 (acute lymphoblastic leukemia), CD30, CD40, CD70, CD133, 57 kDcytokeratin, epithelial specific antigen, epithelial cell adhesionmolecule (EpCAM), extracellular matrix glycoprotein tenascin, Fas/CD95,folate receptor, gastrin-releasing peptide-like receptors, hepatocytespecific antigen, human gastric mucin, human milk fat globule, lymphaticendothelial cell marker, matrix metalloproteinase 9, melan A, melanomamarker, mesothelin, mucin glycoproteins (e.g., MUC1, MUC2, MUC4, MUC5AC,MUC6), prostate specific antigen, prostatic acid phosphatase, PTEN,renal cell carcinoma marker, RGD-peptide binding integrins, sialyl LewisA, six-transmembrane epithelial antigen of the prostate (STEAP), TNFreceptor, TRAIL receptor, tyrosinase, villin. Other tumor associatedantigens include, but are not limited to, alpha fetoprotein,apolipoprotein D, clusterin, chromogranin A, myeloperoxidase, MyoD1myoglobin placental alkaline phosphatase c-fos, homeobox genes.

In an aspect, a detectable label including tumor cell-associatedrecognition components can be used. Many are available from a commercialsource. For example, lectins concanavalin A and wheat germ agglutininare available conjugated to Alexa fluors, Marina Blue, AMCA, OregonGreen, tetramethylrhodamine, Texas Red, fluorescein (from, Invitrogen,Carlsbad, Calif.). Other lectins conjugated to fluorescent dyes areavailable including Phaseolus vulgaris lectin (PHA-L), Arachis hypogaealectin (PNA), Helix pomatia agglutinin (HPA), Soybean agglutinin (SBA),and lectins from Griffonia simplicifolia (from, Invitrogen, Carlsbad,Calif.). Magnetic beads with an antibody to the human epithelialantigen, EpCAM (epithelial cell adhesion molecule) are commerciallyavailable (from ,e.g., Dynal Biotech, Brown Deer, Wis.). EpCAM can beused to selectively bind circulating tumor cells of epithelial origin inthe blood fluid or lymph fluid of a mammalian subject. Anti-CA-125(anti-carcinoembryonic antigen) antibodies can be used to selectivelybind circulating tumor cells of ovarian cancer origin in the blood fluidor lymph fluid of a mammalian subject. Anti-CA125 antibodies can beconjugated to rhodamine-X (Invitrogen, Eugene, Oreg.). Anti-FR(anti-folate receptor) antibodies and folate-FITC, folate-Tc99m can beused to selectively bind circulating tumor cells that overexpress folatereceptors, e.g., ovarian cancer cells, and circulating tumor cells inthe blood fluid or lymph fluid of a mammalian subject. Endocyte, Inc.,West Lafayette, Ind. See, e.g., He, et al., Proc. Natl. Acad. Sci. USA104: 11760-11765, 2007, which is incorporated herein by reference.

In an aspect, the target recognition element is configured to recognizea biomolecule associated with the surface of a pathogen, e.g., bacteria,a virus-infected cell, a fungus, or a parasite. The biomolecule can beone or more components of the bacterial outer membrane, cell wall,and/or cytoplasmic membrane. Examples of components associated with thebacterial outer membrane of Gram-negative bacteria include, but are notlimited to, lipopolysaccaride and OMP (outer membrane protein) porins,the latter of which are exemplified by OmpC, OmpF and PhoP of E. coli.Examples of components associated with the bacterial cell wall of bothGram-positive and Gram-negative bacterial include, but are not limitedto, peptidoglycans polymers composed of an alternating sequence ofN-acetylglucoamine and N-acetyl-muraminic acid and crosslinked by aminoacids and amino acid derivatives. Examples of components associated withthe bacterial cytoplasmic membrane include, but are not limited to, theMPA1-C (also called polysaccharide copolymerase, PCP2a) family ofproteins, the MPA2 family of proteins, and the ABC bacteriocin exporteraccessory protein (BEA) family of proteins. Other examples of componentsassociated with bacteria include, but are not limited to, transporters,e.g., sugar porter (major facilitator superfamily),amino-acid/polyamine/organocation (APC) superfamily, cation diffusionfacilitator, resistance-nodulation-division type transporter, SecDF,calcium:cation antiporter, inorganic phosphate transporter, monovalentcation:proton antiporter-1, monovalent cation:proton antiporter-2,potassium transporter, nucleobase:cation symporter-2, formate-nitritetransporter, divalent anion:sodium symporter, ammonium transporter, andmulti-antimicrobial extrusion; channels, e.g., major intrinsic protein,chloride channel, and metal ion transporter; and primary activetransporters, e.g., P-type ATPase, arsenite-antimonite efflux, Type IIsecretory pathway (SecY), and sodium-transporting carboxylic aciddecarboxylase. A number of other potential components associated withbacteria have been described in Chung, et al., J. Bacteriology183:1012-1021, 2001, which is incorporated herein by reference.

In an aspect, the target recognition element is configured to recognizea biomolecule associated with a blood cell infected with a pathogen. Insome instances, the target recognition element can be a biomoleculeexpressed on the surface of the cell that is derived from the pathogen.For example, red blood cells infected with P. falciparum can bedistinguished from normal red blood cells by changes in surface proteinexpression including expression on the red blood cell surface of theparasite derived protein P. falciparum erythrocyte membrane protein(PfEMP1). See, e.g., Horata, et al., Malaria J. 8:184, 2009, which isincorporated herein by reference.

Controller in Communication with and Responsive to a Sensor The devicecan further include a controller that is in communication with andconfigured to be informed by the one or more sensors. The one or moresensors is operably coupled to the controller, either wirelessly or bycircuit, and can transmit data to the controller regarding the sensedlevels (relative or absolute) of one or more target components in theblood of a vertebrate subject. The controller can be integrated into thedevice. Alternatively, the controller can be a separate component of thedevice that receives and transmits data and/or commands either with orwithout wires. For example, an implanted device can send data regardingthe sensed levels of one or more target components to an externalcontroller through a wireless signal.

The controller can compare the input data regarding the one or moretarget components in the blood fluid or lymph fluid of a vertebratesubject with stored data, or the data can be stored off site and coupledeither wirelessly or by circuit to the sensor and the controller. Thecontroller itself can include the stored data. Alternatively oradditionally, the controller can have access to one or more remotedatabases that include the stored data. The stored data can be dataregarding the normal levels of one or more target components in normalor healthy subjects without a disease, condition, or infection. Thestored data can further include data regarding the baseline level of oneor more target components in a vertebrate subject prior to the onset ofa disease, condition, or infection. The stored data can further includedata regarding the level of one or more target components in avertebrate subject at one or more previous time points. The controllerassesses the most recently obtained input data with the stored data andis configured to controllably initiate steps to release an amount oramounts of one or more bifunctional tags from the one or more firstreservoirs.

The device including the sensor and the controller can also be incommunication with and configured to be informed by one or more tagsensors. The one or more tag sensors can transmit data to the controllerregarding the presence of one or more bifunctional tag/target componentcomplexes. In response to input data, the controller can cause thedevice to controllably divert all or part of the blood fluid or lymphfluid of a vertebrate subject into at least one lumen. Access to the atleast one lumen can be controlled by at least one flow-modulatingelement. A flow-modulating element can be a gate, a valve, a louver, asplitter or flow divider, a filter, a baffle, a channel restriction, aretractable iris, or other structure that controllably limits or permitsaccess of the blood flow to the at least one lumen and the treatmentregion. The controller is operably coupled, either wirelessly or bycircuit, to at least one flow-modulating element. The controller cansend a signal to the at least one flow-modulating element indicatingwhether or not all or part of the flow of blood should be diverted intothe at least one lumen.

The device including the sensor and the controller can also be incommunication with and configured to be informed by one or more bindingsensors. The one or more binding sensors can transmit data to thecontroller regarding the binding of one or more bifunctional tag/targetcomponent complexes to a binding agent within one or more secondreservoirs. In response to input data, the controller can furthercontrollably initiate release or activation of one or more reactivecomponents designed to alter, inactivate or disrupt the one or moretarget components. The one or more reactive components are controllablyreleased or activated in the one or more second reservoirs of thedevice. In an aspect, the controller can release one or more reactivecomponents to modulate the activity of one or more target components.Alternatively or additionally, the controller can send data regardingthe levels of one or more target components in the blood of a vertebratesubject to the subject, to one or more third party individuals such as aphysician or other caregiver, to a computing device, or to a combinationthereof. The subject and/or caregiver or computing device can choose toinitiate steps to alter, inactivate or disrupt the one or more targetcomponents by releasing or activating one or more reactive componentsinto the circulation, into the treatment region, or a combinationthereof.

The device including the controller can also include a processor ornon-volatile memory structure including one or more algorithms residingon the memory that provide computational models of a disease, condition,or infection. For example, a computational model of a disease,condition, or infection can include information regarding, for example,a variety of interrelated signaling pathways involved in the diseaseprocess. The computational model can further inform decisions made bythe controller. Examples of computational models related to inflammatorydisease, cancer and pathogen infection have been described. See, e.g.,U.S. Pat. No. 7,415,359 B2; U.S. Patent Applications 2007/0083333 A1,2008/0201122 A1; Vodovotz, et al., Curr. Opin. Crit. Care. 10:383-390,2004; Zenker, et al., PLoS Comput. Biol. 3(11):e204, 2007; Li, et al.,PLoS ONE 3(7):e2789, 2008; Vodovotz, et al., PLoS Comput. Biol.4:e1000014, 2008; An, Theoretical Biology Medical Modeling 5:11, 2008;Lee, et al., Proc. Natl. Acad. Sci. USA. 104:13086-13091, 2007; Zhou, etal., HIV Medicine. 6:216-223, 2005, each of which is incorporated hereinby reference.

Bifunctional Tags

The device includes one or more bifunctional tags stored in and releasedfrom one or more first reservoirs. The bifunctional tags are configuredto bind one or more target components in the blood and/or lymph of avertebrate subject and sequester the one or more target components inone or more second reservoirs associated with the device. Eachbifunctional tag is configured to include at least one first structuralelement and at least one second structural element. The first and secondstructural elements can include one or more binding moiety configured tobind at least one of a target component and at least one of a bindingagent associated with the device. In an aspect, the first and secondstructural elements can be any one of a detection marker, a toxin, atherapeutic agent. The one or more bifunctional tags can further includeadditional structural elements that are any one of an additional bindingmoiety, detection marker, toxin, therapeutic agent, or combinationthereof. The bifunctional tag can be further armed with a cytotoxicmechanism exemplified by radioisotopes, bacterial toxins, inflammatorycytokines, chemotherapeutics, or prodrugs.

In an aspect, the first structural element and the second structuralelement are one or more of a binding moiety configured to bind at leastone of a target component and at least one of a binding agent associatedwith the device. Examples of binding moieties include, but are notlimited to, antibodies, antibody fragments, peptides, oligonucleotides,DNA, RNA, aptamers, protein nucleic acids, proteins, viruses, enzymes,receptors, bacteria, cells, cell fragments, inorganic molecules, organicmolecules, artificial binding substrates formed by molecular imprintingor combinations thereof. The one or more binding moieties of the firststructural element of the bifunctional tag and the one or more bindingmoieties of the second structural element of the bifunctional tag can befrom the same class of binding moieties. For example, the bifunctionaltag can have a first structural element binding moiety that is a firstantibody and a second structural element binding moiety that is a secondantibody wherein the first antibody binds a target component and thesecond antibody binds a binding agent associated with the device.Alternatively, the binding moiety of the first structural element of thebifunctional tag and the binding moiety of the second structural elementcan be from different classes of binding moieties. For example, thebifunctional tag can have a first structural element binding moiety thatis an antibody and a second structural element binding moiety that is anaptamer wherein the antibody binds a target component and the aptamerbinds a binding agent associated with the device, or vise versa.

The one or more binding moieties of the first or second structuralelement of the bifunctional tag can include one or more antibodies thatbind one or more target components. Antibodies or fragments thereof foruse as one or more binding moieties include, but are not limited to,monoclonal antibodies, polyclonal antibodies, Fab fragments ofmonoclonal antibodies, Fab fragments of polyclonal antibodies, Fab₂fragments of monoclonal antibodies, and Fab₂ fragments of polyclonalantibodies, chimeric antibodies, non-human antibodies, fully humanantibodies, among others. Single chain or multiple chainantigen-recognition sites can be used. Multiple chainantigen-recognition sites can be fused or unfused. Antibodies orfragments thereof can be generated using standard methods. See, e.g.,Harlow & Lane (Antibodies: A Laboratory Manual, Cold Spring HarborLaboratory Press; 1^(St) edition 1988), which is incorporated herein byreference. Alternatively, an antibody or fragment thereof directedagainst one or more target component can be generated, for example,using phage display technology. See, e.g., Kupper, et al. BMCBiotechnology 5:4, 2005, which is incorporated herein by reference. Anantibody, a fragment thereof, or an artificial antibody, e.g., Affibody®artificial antibodies (Affibody AB, Bromma, Sweden) can be preparedusing in silico design (Knappik et al., J. Mol. Biol. 296: 57-86, 2000,which is incorporated herein by reference. In an aspect, antibodiesdirected against one or more target components may be available from acommercial source (from, e.g., Novus Biological, Littleton, Colo.;Sigma-Aldrich, St. Louis, Mo.; United States Biological, Swampscott,Mass.).

The one or more binding moieties of the first or second structuralelement of the bifunctional tag can include one or more aptamers thatbind one or more target components. The aptamer can be anoligonucleotide RNA- or DNA-based aptamer. Aptamers are artificialoligonucleotides (DNA or RNA) that can bind to a wide variety ofentities (e.g., metal ions, small organic molecules, proteins, andcells) with high selectivity, specificity, and affinity. Aptamers can beisolated from a large library of 10¹⁴ to 10¹⁵ random oligonucleotidesequences using an iterative in vitro selection procedure often termed“systematic evolution of ligands by exponential enrichment” (SELEX).See, e.g., Cao, et al., Current Proteomics 2:31-40, 2005; Proske, etal., Appl. Microbiol. Biotechnol. 69:367-374, 2005; Jayasena Clin. Chem.45:1628-1650, 1999, each of which is incorporated herein by reference.In general, SELEX can be used to generate aptamers against any of anumber of target components including but not limited to inflammatorymediators, cancer cells, and bacteria. See, e.g., Guthrie, et al.,Methods 38:324-330, 2006; Shangguan, et al., Proc. Natl. Acad. Sci. USA.103:11838-11843; Chen, et al., Biochem. Biophys. Res. Commun.357:743-748, 2007, each of which is incorporated herein by reference.

The one or more binding moieties of the first or second structuralelement of the bifunctional tag can include one or more peptide basedaptamers that bind one or more target components. Peptide aptamers areartificial proteins in which inserted peptides are expressed as part ofthe primary sequence of a structurally stable protein. See, e.g.,Crawford, et al., Brief. Funct. Genomic Proteomic 2:72-79, 2003, whichis incorporated herein by reference. Peptide aptamers can be generatedby screening a target component against yeast two-hybrid libraries,yeast expression libraries, bacterial expression libraries and/orretroviral libraries. Peptide aptamers can have binding affinitiescomparable to antibodies.

The one or more binding moieties of the first or second structuralelement of the bifunctional tag can include one or more peptide receptorligands that bind one or more target components. Examples of peptidereceptor ligands include, but are not limited to, neuropeptides, forexample, enkephalins, neuropeptide Y, somatostatin,corticotropin-releasing hormone, gonadotropin-releasing hormone,adrenocorticotropic hormone, melanocyte-stimulating hormones,bradykinins, tachykinins, cholecystokinin, vasoactive intestinal peptide(VIP), substance P, neurotensin, vasopressin, and calcitonin; cytokines,for example, interleukins (e.g., IL-1 through IL-35), erythropoietin,thrombopoietin, interferon (IFN), granulocyte monocytecolony-stimulating factor (GM-CSF), tumor necrosis factor (TNF), andothers; chemokines, e.g., RANTES, TARC, MIP-1, MCP, and others; growthfactors, for example, platelet derived growth factor (PDGF),transforming growth factor beta (TGFβ), nerve growth factor (NGF),epidermal growth factor (EGF), insulin-like growth factor (IGF), basicfibroblast growth factor (bFGF); other peptide hormones, for example,atrial natriuretic factor, insulin, glucagon, angiotensin, prolactin,oxyocin, and others.

The one or more binding moieties of the first or second structuralelement of the bifunctional tag can include one or more novel peptidesthat bind one or more target components. Novel peptides that bindselective targets can be generated, for example, using phage displaymethodologies. See, e.g., Spear, et al., Cancer Gene Ther. 8:506-511,2001, which is incorporated herein by reference. In this aspect, thephage express novel peptides on the surface as fusion proteins inassociation with a phage major or minor coat protein and can be screenedfor binding interaction with one or more target components.

The one or more binding moieties of the first or second structuralelement of the bifunctional tag can include one or more receptors thatbind one or more target components. All or part of a receptor can beused as a binding moiety. Examples of receptors include but are notlimited to acetylcholine receptors, adenosine receptors, adrenoceptros,GABA receptors, angiotensin receptors, cannabinoid receptors,cholecystokinin receptors, dopamine receptors, glucagon receptors,glucocorticoid receptors, glutamate receptors, histamine receptors,mineralocorticoid receptors, olfactory receptors, opioid receptors,purinergic receptors, secretin receptors, serotonin receptors,somatostatin receptors, steroid hormone receptors, calcium-sensingreceptor, hormone receptors, erythropoietin receptor, and natriureticpeptide receptors. Other examples include type I cytokine receptors suchas type 1 interleukin receptors, erythropoietin receptor, GM-CSFreceptor, G-CSF receptor, growth hormone receptor, oncostatin Mreceptor, leukemia inhibitory factor receptor; type II cytokinereceptors such as type II interleukin receptors, interferon-α/βreceptors, interferon-γ receptor; many members of the immunoglobulinsuperfamily such as interleukin-1 receptor, CSF1, c-kit receptor,interleukin-18 receptor; tumor necrosis factor (TNF) receptor familysuch as TNF receptor 1 (TNF-R1), TNF receptor 2 (TNF-R2), CD27, CD40,and lymphotoxin (3 receptor; chemokine receptors including serpentineCCR and CXCR receptors, such as CCR1 and CXCR4, and interleukin-8receptor; TGF β receptors such as TGF β receptor 1 and TGF β receptor 2.See Ozaki and Leonard, J. Biol. Chem. 277:29355-29358, 2002, which isincorporated herein by reference

In some instances, binding moieties of the first or second structuralelement of the bifunctional tag can include one or more cellularreceptors that recognize and/or bind to bacteria. For example, CD14,which is normally associated with monocyte/macrophages is known to bindlipopolysaccharide associated with gram negative bacteria as well aslipoteichoic acid associated with the gram positive bacteria Bacillussubtilis (see, e.g., Fan, et al. (1999) Infect. Immun. 67: 2964-2968).Other examples of cellular receptors include, but are not limited to,adenylate cyclase (Bordatella pertussis), Gal alpha 1-4Gal-containingisoreceptors (E. coli), glycoconjugate receptors (enteric bacteria),Lewis(b) blood group antigen receptor (Heliobacter pylori), CR3receptor, protein kinase receptor, galactoseN-acetylgalactosamine-inhibitable lectin receptor, and chemokinereceptor (Legionella), annexin I (Leishmania mexicana), ActA protein(Listeria monocytogenes), meningococcal virulence associated Opareceptors (Meningococcus), alpha5beta3 integrin (Mycobacterium avium-M),heparin sulphate proteoglycan receptor, CD66 receptor, integrinreceptor, membrane cofactor protein, CD46, GM1, GM2, GM3, and CD3(Neisseria gonorrhoeae), KDEL receptor (Pseudomonas), epidermal growthfactor receptor (Samonella typhiurium), alpha5beta1 integrin (Shigella),and nonglycosylated J774 receptor (Streptococci) (see, e.g., U.S. PatentApplication 2004/0033584 A1).

The one or more binding moieties of the first or second structuralelement of the bifunctional tag can include one or more lectins thatbind one or more target components. The term “lectin” was originallyused to define agglutinins which could discriminate among types of redblood cells and cause agglutination. Currently, the term “lectin” isused more generally and includes sugar-binding proteins from manysources regardless of their ability to agglutinate cells. Lectins havebeen found in plants, viruses, microorganisms and animals. Because ofthe specificity that each lectin has toward a particular carbohydratestructure, even oligosaccharides with identical sugar compositions canbe distinguished or separated. Some lectins will bind only to structureswith mannose or glucose residues, while others can recognize onlygalactose residues. Some lectins require that the particular sugar is ina terminal non-reducing position in the oligosaccharide, while otherscan bind to sugars within the oligosaccharide chain. Some lectins do notdiscriminate between a and b anomers, while others require not only thecorrect anomeric structure but a specific sequence of sugars forbinding. Examples of lectins include, but are not limited to, algallectins, e.g., b-prism lectin; animal lectins, e.g., tachylectin-2,C-type lectins, C-type lectin-like, calnexin-calreticulin, capsidprotein, chitin-binding protein, ficolins, fucolectin, H-type lectins,I-type lectins, sialoadhesin, siglec-5, siglec-7, micronemal protein,P-type lectins, pentrxin, b-trefoil, galectins, congerins, selenocosmiahuwena lectin-I, Hcgp-39, Ym1; bacterial lectins, e.g., PseudomonasPA-IL, Burkholderia lectins, chromobacterium CV-IIL, Pseudomonas PA IIL,Ralsonia RS-ILL, ADP-ribosylating toxin, Ralstonia lectin, Clostridiumhemagglutinin, botulinum toxin, tetanus toxin, cyanobacterial lectins,FimH, GafD, PapG, Staphylococcal enterotoxin B, toxin SSL11, toxin SSL5;fungal and yeast lectins, e.g., Aleuria aurantia lectin, integrin-likelectin, Agaricus lectin, Sclerotium lectin, Xerocomus lectin, Laetiporuslectin, Marasmius oreades agglutinin, agrocybe galectin, coprinusgalectin-2, Ig-like lectins, L-type lectins; plant lectins, e.g.,alpha-D-mannose-specific plant lectins, amaranthus antimicrobialpeptide, hevein, pokeweed lectin, Urtica dioica UD, wheat germ WGA-1,WGA-2, WGA-3, artocarpin, artocarpus hirsute AHL, banana lectin,Calsepa, heltuba, jacalin, Madura pomifera MPA, MornigaM, Parkialectins, abrin-a, abrus agglutinin, amaranthin, castor bean ricin B,ebulin, mistletoe lectin, TKL-1, cyanovirin-N homolog, and variouslegume lectins; and viral lectins, e.g., capsid protein, coat protein,fiber knob, hemagglutinin, and tailspike protein (see, e.g., E. Bettler,R. Loris, A. Imberty “3D-Lectin database: A web site for images andstructural information on lectins” 3rd Electronic GlycoscienceConference, The interne and World Wide Web, 6-17 October 1997;http://www.cermay.cnrs.fr/lectines/

The one or more binding moieties of the first or second structuralelement of the bifunctional tag can include one or more artificialbinding substrates formed by the process of molecular imprinting. In theprocess of molecular imprinting, a template, e.g., target component, iscombined with functional monomers which upon cross-linking form apolymer matrix that surrounds the template. See Alexander, et al., J.Mol. Recognit. 19:106-180, 2006, which is incorporated herein byreference. Removal of the template leaves a stable cavity in the polymermatrix that is complementary in size and shape to the template. In anaspect, functional monomers of acrylamide and ethylene glycoldimethacrylate can be mixed with one or more target components in thepresence of a photoinitiator and ultraviolet irradiation used tocross-link the monomers. The resulting polymer can be crushed or groundinto smaller pieces and washed to remove the one or more targetcomponents, leaving a particulate matrix material capable of binding oneor more target components. Examples of other functional monomers,cross-linkers and initiators can be used to generate an artificialbinding substrate are provided. See, e.g., U.S. Pat. No. 7,319,038;Alexander, et al., J. Mol. Recognit. 19:106-180, 2006, each of which isincorporated herein by reference. In a further aspect, hydrogels can beused for molecular imprinting. See, e.g., Byrne et al., “Molecularimprinting within hydrogels”, Advanced Drug Delivery Reviews, 54:149-161, 2002, which is incorporated herein by reference. Other examplesof synthetic binders are provided. See, e.g., U.S. Pat. Nos. 6,255,461;5,804,563; 6,797,522; 6,670,427; and 5,831,012; and U.S. PatentApplication 20040018508; and Ye and Haupt, Anal Bioanal Chem. 378:1887-1897, 2004; Peppas and Huang, Pharm Res. 19: 578-587 2002, each ofwhich is incorporated herein by reference.

A bifunctional tag configured with two or more structural elements canbe constructed using a variety of methods including but not limited toone or more chemical cross-links, one or more streptavidin/biotininteractions, one or more fusion protein constructs, one or more bindingto a common substrate, or a combination thereof. In an aspect, the twoor more structural elements are directly associated with one anotherthrough chemical cross-linking, non-covalent linking, or synthesis as asingle molecule. In an aspect, the two or more structural elements areindirectly associated. In this instance, the two or more structuralelements are separately attached to a mobile substrate such as, forexample, a bead or other particle-like substrate capable of beingreleased into the blood fluid or lymph fluid of a subject. Theparticle-like substrate can include a bead, a vesicle, a cell, a carbonnanotube, or other similar structures.

A bifunctional tag configured with two or more structural elements canbe constructed by cross-linking the two or more structural elements toone another. For example, two antibodies with different specificitiescan be cross-linked together to form a bifunctional tag using a chemicalcross-linking agent, e.g., N-succinimidyl 3-(2-pyridyldithio)propionate(SPDP). See, e.g., U.S. Pat. No. 5,470,570, which is incorporated hereinby reference. Any of a number of homobifunctional, heterofunctional,and/or photoreactive cross linking agents can be used to cross-link twoor more structural elements of a bifunctional tag. Examples ofhomobifunctional cross linkers include, but are not limited to, primaryamine/primary amine linkers such as BSOCES((bis(2-[succinimidooxy-carbonyloxy]ethyl) sulfone), DMS (dimethylsuberimidate), DMP (dimethyl pimelimidate), DMA (dimethyl adipimidate),DSS (disuccinimidyl suberate), DST (disuccinimidyl tartate), Sulfo DST(sulfodisuccinimidyl tartate), DSP (dithiobis(succinimidyl propionate),DTSSP (3,3′-dithiobis(succinimidyl propionate), EGS (ethylene glycolbis(succinimidyl succinate)) and sulfhydryl/sulfhydryl linkers such asDPDPB (1,4-di-(3′-[2′pyridyldithio]-propionamido) butane). Examples ofheterofunctional cross linkers include, but are not limited to, primaryamine/sulfhydryl linkers such as MBS(m-maleimidobenzoyl-N-hydroxysuccinimide ester), Sulfo MBS(m-maleimidobenzoyl-N-hydroxysulfosuccinimide), GMBS(N-gamma-maleimidobutyryl-oxysuccinimide ester), Sulfo GMBS(N-γ-maleimidobutyryloxysulfosuccinimide ester),EMCS(N-(epsilon-maleimidocaproyloxy) succinimide ester), SulfoEMCS(N-(epsilon-maleimidocaproyloxy) sulfo succinimide), SIAB(N-succinimidyl(4-iodoacetyl)aminobenzoate), SMCC (succinimidyl4-(N-maleimidomethyl)cyclohexane-1-carboxylate), SMPB (succinimidyl4-(rho-maleimidophenyl) butyrate), Sulfo SIAB(N-sulfosuccinimidyl(4-iodoacetyl)aminobenzoate), Sulfo SMCC(sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate),Sulfo SMPB (sulfosuccinimidyl 4-(rho-maleimidophenyl) butyrate), andMAL-PEG-NHS (maleimide PEG N-hydroxysuccinimide ester);sulfhydryl/hydroxyl linkers such as PMPI (N-rho-maleimidophenyl)isocyanate; sulfhydryl/carbohydrate linkers such asEMCH(N-(epsilon-maleimidocaproic acid) hydrazide); and amine/carboxyllinkers such as EDC (1-ethyl-3-(3-dimethylaminopropyl) carbodiimidehydrochloride).

A bifunctional tag configured with two or more structural elements canbe constructed using one or more interactions between biotin and avidin,streptavidin or derivatives thereof. Streptavidin and avidin aremultivalent proteins capable of binding multiple biotin subunits withhigh affinity and as such can be used as linking molecules between twobiotinylated structural elements. An aptamer or antibody structuralelement directed against one or more target components can be linked toa second structural element of a bifunctional tag using astreptavidin-biotin bridge. For example, a biotinylated aptamer can belinked to a biotinylated antibody through a streptavidin bridge. Anaptamer portion of the bifunctional tag can be biotinylated byintroducing a biotinylated nucleotide into the aptamer sequence duringin vitro transcription. An antibody portion of the bifunctional tag canbe biotinylated using an amine reactive biotinylation reagent such as,for example, EZ-Link Sulfo-NHS-SS-Biotin (sulfosuccinimidyl2-(biotinamido)-ethyl-1,3-dithiopropionate; Pierce-Thermo Scientific,Rockford, Ill., USA. See e.g., Jaiswal, et al. Nature Biotech. 21:47-51,2003, which is incorporated herein by reference. The biotinylatedaptamer is reacted with the biotinylated antibody in the presence ofstreptavidin to generate the bifunctional tag. Alternatively, a firststructural element of the bifunctional tag can contain all or part ofthe streptavidin protein for use in binding to a biotin-modified secondstructural element. For example, cDNA sequence encoding all or part ofan antibody or other protein/peptide can be genetically modified tocontain all or part of the streptavidin gene using standard cloningprocedures, resulting in a streptavidin-antibody fusion protein. See,e.g., Koo, et al. Appl. Environ. Microbiol. 64:2497-2502, 1998, which isincorporated herein by reference.

A bifunctional tag configured with two or more structural elements canbe constructed by association of two non-covalently associatedsingle-chain antibodies. In one aspect, protein engineeringmethodologies are used to generate the variable domains of two distinctantibodies (A and B) and are arranged as VHa-VLb (variable heavy chaindomain of antibody A and variable light chain domain of antibody B) onone chain and VHb-VLa (variable heavy chain domain of antibody B andvariable light chain domain of antibody A) on the other chain togenerate diabody structures with two distinct binding affinities. See,e.g., Takemura, et al., Protein Eng. Des. Sel.

13:583-588, 2000, which is incorporated herein by reference. Thebifunctional tags generated from two distinct antibody variable domainscan be further covalently cross-linked with disulfide bonds for addedstability and to enable addition of a third structural element such as aradiolabel or fluorescent tag. See, e.g., Olafsen, et al., Protein Eng.Des. Sel. 17:315-323, 2004, which is incorporated herein by reference. Anumber of other methods have been described for generating antibody-likebiomolecules with two or more distinct binding specificities. See.,e.g., U.S. Pat. No. 4,444,878; U.S. Patent Application 2009/0182127;Kufer, et al., Trends Biotechnol. 22:238-244, 2004; Stork, et. al.,Protein Eng. Des. Sel. 20:569-576, 2007, each of which is incorporatedherein by reference.

A bifunctional tag configured with two or more structural elements canbe constructed by generating a fusion protein. In this instance, cDNAencoding the two or more structural elements are incorporated into anappropriate expression construct to generate a protein expressionproduct that includes amino acid sequence from each structural elementand retains the binding properties of each structural element . In anaspect, the bifunctional tag includes a first structural element that isan antibody and a second structural element that is streptavidin. See,e.g., Koo, et al. Appl. Environ. Microbiol. 64:2497-2502, 1998, which isincorporated herein by reference. In an aspect, the bifunctional tagincludes a first structural element that is an antibody and a secondstructural element that is a receptor ligand. See, e.g., Becker, et al.,Proc. Natl. Acad. Sci. USA. 93:7826-7831, 1996; Challita-Eid, et al., J.Immunol. 160:3419-3426, 1998, each of which is incorporated herein byreference. In a further aspect, the bifunctional tag includes a firststructural element that is a first receptor ligand and a secondstructural element that is a second receptor ligand. See, e.g., U.S.Pat. No. 6,132,992, which is incorporated herein by reference.

Sensing Occupied Versus Unoccupied Bifunctional Tags

The bifunctional tag can be further configured to controllably emit asignal in response to binding a target component. In an aspect, the oneor more bifunctional tags are configured to emit a signal only when atarget component is bound. Alternatively, the one or more bifunctionaltags are configured to emit a first signal in the absence of a boundtarget component and a second signal in the presence of a bound targetcomponent.

In an aspect, the bifunctional tag can be configured such that bindingof the target component to the tag results in a conformational change inthe bifunctional tag that can be measured using fluorescence resonanceenergy transfer (FRET). FRET is a distance-dependent interaction betweenthe electronic excited states of two dye molecules in which excitationis transferred from a donor molecule to an acceptor molecule withoutemission of a photon. In an aspect, interaction of a donor molecule withan acceptor molecule can lead to a shift in the emission wavelengthassociated with excitation of the acceptor molecule. In other aspects,interaction of a donor molecule with an acceptor molecule can lead toquenching of the donor emission. The one or more structural elements ofthe bifunctional tag can include at least one donor molecule and atleast one acceptor molecule. In this configuration, binding of one ormore target components to the bifunctional tag results in aconformational change in the bifunctional tag and results in a change inthe distance between the donor and acceptor molecules and a change inmeasurable fluorescence.

A variety of donor and acceptor fluorophore pairs can be considered forFRET associated with the bifunctional tag including, but not limited to,fluorescein and tetramethylrhodamine; IAEDANS and fluorescein;fluorescein and fluorescein; and BODIPY FL and BODIPY FL. A number ofAlexa Fluor (AF) fluorophores (Molecular Probes-Invitrogen, Carlsbad,Calif., USA) can be paired with other AF fluorophores for use in FRET.Some examples include, but are not limited, to AF 350 with AF 488; AF488 with AF 546, AF 555, AF 568, or AF 647; AF 546 with AF 568, AF 594,or AF 647; AF 555 with AF594 or AF647; AF 568 with AF6456; and AF594with AF 647.

The cyanine dyes Cy3, Cy5, Cy5.5 and Cy7, which emit in the red and farred wavelength range (>550 nm), offer a number of advantages forFRET-based detection systems. Their emission range is such thatbackground fluorescence is often reduced and relatively large distances(>100 Å) can be measured as a result of the high extinction coefficientsand good quantum yields. For example, Cy3, which emits maximally at 570nm and Cy5, which emits at 670 nm, can be used as a donor-acceptor pair.When the Cy3 and Cy5 are not proximal to one another, excitation at 540nm results only in the emission of light by Cy3 at 590 nm. In contrast,when Cy3 and Cy5 are brought into proximity by a conformation change inan aptamer, antibody, or receptor, for example, excitation at 540 nmresults in an emission at 680 nm.

Quenching dyes are used as part of the bifunctional tag to quench thefluorescence of visible light-excited fluorophores. Examples ofquenching dyes include, but are not limited, to DABCYL, thenon-fluorescing diarylrhodamine derivative dyes QSY 7, QSY 9 and QSY 21(Molecular Probes, Carlsbad, Calif., USA), the non-fluorescing BlackHole Quenchers BHQ0, BHQ1, BHQ2, and BHQ3 (Biosearch Technologies, Inc.,Novato, Calif., USA) and Eclipse (Applera Corp., Norwalk, Conn., USA). Avariety of donor fluorophore and quencher pairs can be considered forFRET associated with the bifunctional tag including, but not limited to,fluorescein with DABCYL; EDANS with DABCYL; or fluorescein with QSY 7and QSY 9. In general, QSY 7 and QSY 9 dyes efficiently quench thefluorescence emission of donor dyes including blue-fluorescentcoumarins, green- or orange-fluorescent dyes, and conjugates of theTexas Red and Alexa Fluor 594 dyes. QSY 21 dye efficiently quenches allred-fluorescent dyes. A number of the Alexa Fluor (AF) fluorophores(Molecular Probes-Invitrogen, Carlsbad, Calif., USA) can be paired withquenching molecules as follows: AF 350 with QSY 35 or DABCYL; AF 488with QSY 35, DABCYL, QSY7 or QSY9; AF 546 with QSY 35, DABCYL, QSY7 orQSY9; AF 555 with QSY7 or QSY9; AF 568 with QSY7, QSY9 or QSY21; AF 594with QSY21; and AF 647 with QSY 21.

The bifunctional tag can include at least one structural element that isan RNA or DNA oligonucleotide-based aptamers with one or morefluorescent tags and one or more quenching tags. Upon binding of atarget component, the aptamer undergoes a conformational shift such thatthe distance between the donor fluorophore and the acceptor fluorophoreor quencher is altered, leading to a change in measurable fluorescence.Aptamers can be isolated from a large library of 10¹⁴ to 10¹⁵ randomoligonucleotide sequences using an iterative in vitro selectionprocedure often termed “systematic evolution of ligands by exponentialenrichment” (SELEX). Aptamers can be generated against virtually anyclass of molecules including cells (e.g., cancer cells, bacteria, andparasites), proteins (e.g., hormones), and chemicals (e.g., codeine,cocaine). See, e.g., Shangguan, et al., Proc. Natl. Acad. Sci. USA.103:11838-11843; Chen, et al., Biochem. Biophys. Res. Commun.357:743-748, 2007; Ulrich, et al., J. Biol. Chem. 277:20756-20762, 2002;Cao, et al. Current Proteomics 2:31-40, 2005; Proske, et al. Appl.Microbiol. Biotechnol. 69:367-374, 2005, Win, et al. Nucleic Acids Res.34:5670-5682, 2006, each of which is incorporated herein by reference.For example, an aptamer that selectively binds cocaine can be generatedusing SELEX as described above and modified to incorporate a fluorophoresuch as, for example, fluorescein and a quencher such as, for example,4-[4′-((dimethylamino)phenyl)azo-] benzoic acid (DABCYL). See, e.g.,Stojanovic, et al. J. Am. Chem. Soc. 123:4928-4931, 2001, which isincorporated herein by reference. In this instance, binding of cocaineto the aptamer induces a conformational change in the aptamer thatcauses the fluorophore and the quencher to move closer in proximity. Assuch, the presence of cocaine can be measured as a function of thedecrease in or quenching of the fluorescein emission at a wavelength of518 nm.

Semiconductor quantum dots (QDs) with various excitation/emissionwavelength properties can be used to label an aptamer structural elementof a bifunctional tag. Various methods are available for attachingquantum dots to the DNA backbone of an aptamer such as, for example,covalent linkage of amino-labeled DNA to carboxylated QDs and linkage ofbiotinylated DNA to streptavidin modified QDs. See, e.g., Cady, et al.J. Mol. Cell. Probes 21:116-124, 2007, which is incorporated herein byreference. For example, carboxy QDs (from, e.g., Quantum DotCorporation, Hayward, Calif., USA) can be attached to an aptamer througha C6 amino modifier placed on either the 5 prime or 3 prime end of theaptamer sequence. Alternatively, streptavidin QDs (from, e.g., QuantumDot Corporation, Hayward, Calif., USA) can be attached to an aptamerthrough a biotin attached to the 5-prime end of the aptamer sequence.

The fluorophores can be attached to various chemical moieties that allowfor attachment at various sites within the aptamer. For example,3′-DABCYL CPG can be used to place the fluorophore DABCYL at the 3 primeterminus of the aptamer whereas 5′-DABCYL phosphoramidite can be used toplace DABSYL at the 5 prime terminus of the aptamer (see, e.g., productinformation at Glen Research, Sterling, Va.). DABCYL dT can be used toplace DABCYL within the body of the aptamer sequence. Labeling aptamerswith appropriate commercially available fluorophores can be achievedfollowing instructions provided by the respective manufacturer.Alternatively, custom made aptamer-based molecular beacons are availablefrom commercial sources (from, e.g., Biosearch Technologies, Inc.,Novato, Calif., USA).

In some instances, an aptamer can have a fluorophore in a region of themolecule known to undergo conformational change upon binding of a targetthat leads to an increase in fluorescence intensity. An aptamer of thissort can be selected using an in vitro selection process withfluorescently labeled aptamers. See, e.g., Jhaveri, et al. NatureBiotech. 18:1293-1297, 2000, which is incorporated herein by reference.A pool of RNA molecules is generated in which the random sequence region(45-60 residues) is skewed such that one of the residues, uridine, forexample, is disproportionately underrepresented. The three to fourrandomly placed uridine residues are substituted withfluorescein-12-UTP, Cascade Blue-7-UTP, Texas Red-5-UTP, and/orRhodamine Green-5-UTP during in vitro transcription. The labeled pool ofRNA molecules are screened against a target component by passing thelabeled pool over a column matrix or other matrix to which the targetcomponent is attached. Those RNA molecules that bind with high affinityto the target component are further screened for their fluorescencesignaling properties in response to binding of the target component. Forexample, the baseline fluorescence intensity is measured for RNA aptamermolecules labeled with fluorescein-12-UTP (excitation maxima 494 nm,emission maxima 521 nm) or Rhodamine Green-5-UTP (excitation maxima 505nm, emission maxima 533 nm), for example, then re-measured in responseto increasing concentrations of the target component. Fluorescentaptamers can be selected that exhibit a 100-200% increase influorescence intensity in response to target binding. See, e.g.,Jhaveri, et al. Nature Biotech. 18:1293-1297, 2000, which isincorporated herein by reference.

In an aspect, the bifunctional tag can include at least one structuralelement that is an antibody with one or more donor fluorophore and oneor more acceptor fluorophore or quencher. Upon binding of a targetcomponent, the antibody undergoes a conformational shift such that thedistance between the donor fluorophore and the acceptor fluorophore orquencher is altered, leading to a change in measurable fluorescence. Theantibody can be designed to emit a shift in emission wavelength, forexample, in response to binding a target component. An antibody thatexhibits a shift in fluorescent signal in response to binding of anantigen can be generated by labeling the antibody with asolvent-sensitive fluorophore, e.g., dansyl chloride(5-dimethylaminonaphthalene-1-sulfonyl chloride). See, e.g., Brennan J.Fluor. 9:295-312, 1999, which is incorporated herein by reference. Theantibody is labeled such that binding of the target component to theantibody shields the solvent sensitive fluorescent label near the activebinding site from the solvent water, resulting in a 3-5 fold increase influorescence intensity. See, e.g., Bright, et al. Anal. Chem.62:1065-1069, 1990, which is incorporated herein by reference.

In an aspect, the bifunctional tag can include at least one structuralelement that is an antibody that signals binding of a target componentusing FRET and a flexible arm. For example, an antibody can include adonor fluorophore near the binding site or the target component as wellas a flexible arm containing an analog of the target component that islabeled with a quencher and recognized by the antibody. See, e.g. U.S.Patent Application 2006/0172318 A1, which is incorporated herein byreference. As the labeled target component analog moves into proximityto the labeled active site, a baseline FRET signal can be measured. Ameasurable change in the FRET signal is detected when the analog iscompetitively displaced by the actual target component. The flexible armcan be composed of DNA, RNA, polymers, protein nucleic acid (PNA),peptides, protein or oligosaccharide. For example, an aminofunctionalized DNA arm can be treated with a bifunctional NHS-esteractivated Cy3.5 dye to add a fluorescent tag to the flexible arm. Theanalog of the target component is modified with a monoamine andinteracted with the bifunctional NHS-ester and attached to the DNAflexible arm. See, e.g., U.S. Patent Application 2006/0172318 A1, whichis incorporated herein by reference. The flexible arm can be attacheddirectly to the antibody through a thiol-maleimide linkage such that theDNA flexible arm is modified with a thiol group at one end and linkedvia maleimide to one or more cysteine groups on the antibody.Alternatively, the flexible arm can be attached to a protein, forexample, that is adjacent to the antibody or to which the antibody isbound.

The device can periodically emit electromagnetic energy sufficient toexcite the fluorophores associated with, for example, an aptamer orantibody on a bifunctional tag and as such measure emitted fluorescence.The excitation and emission wavelengths used can range from 320 nm to1000 nm, depending upon the fluorophores used. For example, Cy5 isexcited at 649 nm and has a maximum emission at 670 nm. The light sourcecan be a broad band white light source filtered to provide theappropriate excitation wavelength. Alternatively, the light source canbe a laser such as, for example, gas lasers, eximer lasers, solid statelasers, and dye lasers. In some instances, one or more laser diodes, forexample, can be incorporated into the device. A charge coupled device(CCD) or other light capturing device can be used to detect the emittedfluorescence.

First Reservoirs for Storing and Releasing Bifunctional Tags

The device includes one or more first reservoirs configured to store andcontrollably release one or more bifunctional tags. In an aspect, thedevice includes a single first reservoir configured to store one or morebifunctional tags. In an aspect, the device includes multiple firstreservoirs wherein each multiple first reservoir is configured to storeone or more bifunctional tags. The one or more first reservoirs can bean integral part of the device. Alternatively, the one or more firstreservoirs can be a separate part of the device, located proximal to orat a distance from the main body of the device, but in wirelesscommunication with the main body of the device.

Release of one or more bifunctional tags from the one or more firstreservoirs can be facilitated by one or more release mechanisms. In anaspect, the bifunctional tags can be continuously released from at leastone first reservoir at a constant rate over time. For example, therelease mechanism can use one or more of a slow release, controlledrelease, or extended release biodegradable composition that dissolves orbreaks down over time. Examples of slow release, controlled release, orextended release compositions include but are not limited to hydrogels,polymers, gelled and/or cross-linked water swellable polyolefins,polycarbonates, polyesters, polyamides, polyethers, polyepoxides andpolyurethanes such as, for example, poly(acrylamide), poly(-hydroxyethylacrylate), poly(2-hydroxypropyl acrylate), poly(N-vinyl-2-pyrrolidone),poly(n-methylol acrylamide), poly(diacetone acrylamide),poly(-hydroxylethyl methacrylate), poly(allyl alcohol). Other suitablepolymers include but are not limited to cellulose ethers, methylcellulose ethers, cellulose and hydroxylated cellulose, methyl celluloseand hydroxylated methyl cellulose, gums such as guar, locust, karaya,xanthan gelatin, and derivatives thereof. The rate of dissolution of thecomposition containing the bifunctional tag can be monitored using animpedance-based sensor as described by Johnson et al., in J. ElectrochemSoc. 152:H6-H11, 2005, which is incorporated herein by reference.

In an aspect, the one or more bifunctional tags can be controllablyreleased from the one or more first reservoirs and can include periodsof release followed by periods of non-release. Controlled release fromone or more first reservoirs can include a release mechanism thatreversibly opens and closes a portion of the first reservoir. Therelease mechanism can include a variety of different types of releasemechanisms, including, for example, a controllable valve. Variousexamples of micro valves or microelectromechanical systems (MEMS) valvesfor controlling fluid flow have been described. See, e.g., Luckevich M.Valve World, May 2007, pp. 79-83; Givrad TK., et al., Proceedings ofBIOMed2008, 3^(rd) Frontiers in Biomedical Devices Conference. Jun.18-20, 2008, Irvine, Calif., USA; U.S. Pat. Nos. 6,612,535; 7,124,773,each of which is incorporated herein by reference.

In an aspect, the one or more first reservoirs are covered with amaterial that can be controllably removed or punctured to release one ormore bifunctional tags. The cover material can be responsive to adirectly applied stimulus (e.g., an applied voltage or potential) or toa change in the local environment of the device (e.g., local pH change),or any of a number of other stimuli including but not limited to heat,light (e.g., laser), and magnetic field. See, e.g., U.S. Pat. No.6,808,522; Grayson, R. et al., Proceedings of IEEE 92:6-21, 2004, whichare each incorporated herein by reference. As an example, the one ormore first reservoirs can be an array of microreservoirs on a microchipin which each aliquot of one or more bifunctional tags is contained inits own reservoir and capped by an environmentally sensitive material.In one aspect, the microreservoirs can be capped with a gold membranewhich is weakened and ruptured by electrochemical dissolution inresponse to application of an anode voltage to the membrane in thepresence of chloride ions, resulting in release of drug as described inU.S. Pat. No. 5,797,898 and in Prescott, et al., Nat. Biotech.,24:437-438, 2006, each of which is incorporated herein by reference.Alternatively, the microreservoirs can be capped by a temperaturesensitive material which can be ruptured in response to selectiveapplication of heat to one or more of the reservoirs as described inU.S. Pat. No. 6,669,683, which is incorporated herein by reference.Wireless induction of a voltage or thermal trigger, for example, to agiven reservoir of a microarray of reservoirs by a subject would enableon-demand release of one or more bifunctional tags.

In some instances, the one or more first reservoirs of the device canincorporate a natural and/or synthetic stimulus-responsive hydrogel orpolymer which changes confirmation rapidly and reversibly in response toenvironmental stimuli such as, for example, temperature, pH, ionicstrength, electrical potential, light, magnetic field or ultrasound. Seee.g., Stubbe, et al., Pharmaceutical Res., 21:1732-1740, 2004, which isincorporated herein by reference. Examples of polymers are described inU.S. Pat. Nos. 5,830,207; 6,720,402; and 7,033,571, each of which isincorporated herein by reference. In some instances, the one or morebifunctional tags can be dissolved or dispersed in the hydrogel orpolymer. Alternatively, a hydrogel and/or other stimulus-responsivepolymer can be incorporated into the release mechanism. For example, ahydrogel or other polymer or other smart material can be used as anenvironmentally sensitive actuator to control flow of a therapeuticagent out of an implantable device as described in U.S. Pat. Nos.6,416,495; 6,571,125; and 6,755,621, each of which is incorporatedherein by reference. The one or more first reservoirs can incorporate ahydrogel or other polymer that modulates delivery of one or morebifunctional tags in response to a trigger from the sensor-informedcontroller.

In some instances, release of one or more bifunctional tags from thefirst reservoirs can be nonprogammable, delivered as a predetermineddosage. For example, the one or more bifunctional tags can beadministered using continuous infusion. Alternatively, the release ofone or more bifunctional tag from the first reservoirs can be linked toa timing mechanism associated with the device. For example, the timingmechanism can instruct release of one or more bifunctional tags at agiven time of day, a given time of week, a given time of month, and/or agiven time of year. The timing mechanism can be further linked to inputfrom the one or more sensors, releasing one or more bifunctional tag atone or more predetermined time following detection of one or more targetcomponents.

The release of one or more bifunctional tags from the first reservoirscan be programmable, having on/off and/or variable delivery rates basedon either external or internal control. External control can be mediatedby manual manipulation of a hand-operated pulsative pump with one-wayvalves associated with a device implanted near the surface of the skin,for example. Alternatively, external control can be mediated by remotecontrol through an electromagnetic wireless signal such as, for example,infrared or radio waves that are able to trigger an electrical stimuluswithin the implanted device. Examples of remote control drug deliverydevices are described in U.S. Pat. Nos. 5,928,195; 6,454,759; and6,551,235, each of which is incorporated herein by reference. As such,one or more bifunctional tag can be delivered by continuous infusion inresponse to an “on” trigger and stopped in response to an “off” trigger,for example. Alternatively, one or more bifunctional tag can bedelivered as a microbolus, for example, in response to an “on” triggeras described in U.S. Pat. No. 6,554,822, which is incorporated herein byreference. External control can be initiated by a caregiver.Alternatively, a subject can initiate delivery of one or morebifunctional tag. As such, the system can have a built in mechanism tolimit the number of allowable doses by a subject and/or caregiver in agiven time frame as described, for example, in U.S. Pat. No. 6,796,956,which is incorporated herein by reference.

Binding of Bifunctional Tag in Second Reservoir

The device disclosed herein further includes one or more secondreservoirs. The one or more second reservoirs are configured to bind andsequester one or more target components by selectively capturingbifunctional tags to which are bound one or more target components. Theone or more second reservoirs can include one or more reactivecomponents that are binding agents configured to bind one or morestructural elements of a bifunctional tag. The one or more bindingagents can be used alone to selectively or non-selectively sequester oneor more bifunctional tags. Alternatively, the one or more binding agentscan be used to capture one or more bifunctional tag in combination withtreatment including one or more additional reactive components, e.g., adenaturing agent, a degradative agent, a cell-disrupting agent, amodulator, an energy source, an apoptotic agent, a programmed celldeath-inducing agent, a cytotoxic agent, cytostatic agent, achemotherapeutic agent, an antibody toxin or a combination thereof.Following binding of the one or more bifunctional tags to the one ormore binding agents in a second reservoir, one or more additionalreactive components can be provided to inactive the one or more targetcomponents bound to the bifunctional tag.

The one or more binding agents can include absorbent or adsorbentmaterial that non-selectively binds one or more bifunctional tags. Theabsorbent or adsorbent material can include, but is not limited to,silica, activated charcoal, nonionic or uncharged resins or polymers,ionic or charged resins or polymers, anion exchange resins or polymers,cation exchange resins or polymers, neutral exchange resins or polymers,immobilized polymyxin B, immobilized monoclonal antibodies, immobilizedinflammatory mediator receptors, immobilized specific antagonists,cellulose, cellulose derivatives, synthetic materials, polysulfone,polyacrylonitrile, polymethylmethacrylate, polyvinyl-alcohol, polyamide,polycarbonate, polystyrene-derivative fibers, and any combinationthereof. Specific examples of absorbent or adsorbent materials that havebeen used in animal and clinical studies for non-specific binding ofinflammatory mediators, for example, include, but are not limited to,polystyrene-divinylbenzene copolymer beads with biocompatiblepolyvinylpyrrolidone coating (CYTOSORB, MedaSorb Corporation, NJ, USA)and 2-methacryloyloxyethyl phosphorylcholine (MPCF-X; see, e.g., Nakada,et al., Transfus. Apher. Sci. 35:253-264, 2006, which is incorporatedherein by reference.

In an aspect, the one or more binding agents can include one or morebiomolecules that non-specifically bind immunoglobulins, examples ofwhich include anti-immunoglobulin antibodies, Protein A and Protein G.In a further aspect, the binding agent can be avidin or streptavidin forbinding biotinylated bifunctional tags. Alternatively, the binding agentcan be biotin for binding avidin or streptavidin labeled bifunctionaltags.

The one or more binding agents can selectively bind one or morestructural elements of the bifunctional tag. A selective binding agentcan include, but is not limited to, an antibody or fragments thereof, anoligonucleotide or peptide based aptamer, a receptor or parts thereof, aligand, an artificial binding substrate formed by molecular imprinting,or combinations thereof. The reactive components can further include oneor more of an adhesion molecule, a binding mimetic, a polymer, a lectin,integrin, or selectin. In some instances, the one or more structuralelements of the bifunctional tag is itself an antibody or fragmentsthereof, an oligonucleotide or peptide based aptamer, a receptor orparts thereof, a ligand, an artificial binding substrate formed bymolecular imprinting, or combinations thereof, in which case, thebinding agent can be an antigen, an epitope, a binding partner, areceptor, a ligand, and the like.

The one or more binding agents can include one or more antibodies thatbind one or more structural elements of the bifunctional tag. Antibodiesor fragments thereof for use as one or more binding agents include, butare not limited to, monoclonal antibodies, polyclonal antibodies, Fabfragments of monoclonal antibodies, Fab fragments of polyclonalantibodies, Fab₂ fragments of monoclonal antibodies, and Fab₂ fragmentsof polyclonal antibodies, chimeric antibodies, non-human antibodies,fully human antibodies, among others. Single chain or multiple chainantigen-recognition sites can be used. Multiple chainantigen-recognition sites can be fused or unfused. Antibodies orfragments thereof can be generated using standard methods. See, e.g.,Harlow & Lane (Antibodies: A Laboratory Manual, Cold Spring HarborLaboratory Press; 1^(st) edition 1988), which is incorporated herein byreference. Alternatively, an antibody or fragment thereof directedagainst one or more inflammatory mediators can be generated, forexample, using phage display technology. See, e.g., Kupper, et al. BMCBiotechnology 5:4, 2005, which is incorporated herein by reference. Anantibody, a fragment thereof, or an artificial antibody, e.g., Affibody®artificial antibodies (Affibody AB, Bromma, Sweden) can be preparedusing in silico design (Knappik et al., J. Mol. Biol. 296: 57-86, 2000,which is incorporated herein by reference.

The one or more binding agents can include one or more aptamers thatbind one or more structural elements of the bifunctional tag. Theaptamer can be an oligonucleotide RNA- or DNA-based aptamer. Aptamersare artificial oligonucleotides (DNA or RNA) that can bind to a widevariety of entities (e.g., metal ions, small organic molecules,proteins, and cells) with high selectivity, specificity, and affinity.Aptamers can be isolated from a large library of 10¹⁴ to 10¹⁵ randomoligonucleotide sequences using an iterative in vitro selectionprocedure often termed “systematic evolution of ligands by exponentialenrichment” (SELEX). See, e.g., Cao, et al., Current Proteomics 2:31-40,2005; Proske, et al., Appl. Microbiol. Biotechnol. 69:367-374, 2005;Jayasena Clin. Chem. 45:1628-1650, 1999, each of which is incorporatedherein by reference.

In an aspect, the one or more binding agents can include one or moreaptamers that are peptide based aptamers. Peptide aptamers areartificial proteins in which inserted peptides are expressed as part ofthe primary sequence of a structurally stable protein. See, e.g.,Crawford, et al., Brief Funct. Genomic Proteomic 2:72-79, 2003, which isincorporated herein by reference. Peptide aptamers can be generated byscreening the one or more structural elements of the bifunctional tagagainst yeast two-hybrid libraries, yeast expression libraries,bacterial expression libraries and/or retroviral libraries. Peptideaptamers can have binding affinities comparable to antibodies.

In a further aspect, the one or more binding agents can include one ormore novel peptides. Novel peptides that bind selective targets can begenerated, for example, using phage display methodologies. See, e.g.,Spear, et al., Cancer Gene Ther. 8:506-511, 2001, which is incorporatedherein by reference. In this aspect, the phage express novel peptides onthe surface as fusion proteins in association with a phage major orminor coat protein and can be screened for binding interaction with oneor more structural element of the bifunctional tag.

The one or more binding agents can include one or more artificialbinding substrates formed by the process of molecular imprinting andconfigured to bind one or more structural elements of the bifunctionaltag. In the process of molecular imprinting, a template is combined withfunctional monomers which upon cross-linking form a polymer matrix thatsurrounds the template. See Alexander, et al., J. Mol. Recognit.19:106-180, 2006, which is incorporated herein by reference. Removal ofthe template leaves a stable cavity in the polymer matrix that iscomplementary in size and shape to the template. In an aspect,functional monomers of acrylamide and ethylene glycol dimethacrylate canbe mixed with structural elements of the bifunctional tag in thepresence of a photoinitiator and ultraviolet irradiation used tocross-link the monomers. The resulting polymer can be crushed or groundinto smaller pieces and washed to remove the structural elements of thebifunctional tag, leaving a particulate matrix material capable ofbinding one or more structural elements of the bifunctional tag.Examples of other functional monomers, cross-linkers and initiators canbe used to generate an artificial binding substrate are provided. See,e.g., U.S. Pat. No. 7,319,038; Alexander, et al., J. Mol. Recognit.19:106-180, 2006, each of which is incorporated herein by reference. Ina further aspect, hydrogels can be used for molecular imprinting. See,e.g., Byrne et al., “Molecular imprinting within hydrogels”, AdvancedDrug Delivery Reviews, 54: 149-161, 2002, which is incorporated hereinby reference. Other examples of synthetic binders are provided. See,e.g., U.S. Pat. Nos. 6,255,461; 5,804,563; 6,797,522; 6,670,427; and5,831,012; and U.S. Patent Application 20040018508; and Ye and Haupt,Anal Bioanal Chem. 378: 1887-1897, 2004; Peppas and Huang, Pharm Res.19: 578-587 2002, each of which is incorporated herein by reference.

In an aspect, the one or more binding agents can be configured topreferentially bind a bifunctional tag that is bound to or occupied byan appropriate target component. The binding agents are furtherconfigured such that unbound or unoccupied bifunctional tags and/ortarget components are unable to bind to the binding agents and remain inthe blood or lymph circulation. Binding agents configured to bind onlythe bifunctional tag/target component complex can be generated bydeveloping screening assays for binding agents that preferentially bindthe complex. For example, an antibody screen can be developed using aphage display library and a bifunctional tag/target component complex asthe screening antigen. Once antibodies are found that bind thebifunctional tag/target component complex, additional negative screensare performed to eliminate those antibodies that bind either thebifunctional tag alone or the target component alone. See, e.g., Nemazee& Sato, Proc. Natl. Acad. Sci, USA. 79:3828-3832, 1982; Malek et al.,Proc. Natl. Acad. Sci., USA. 80:5694-5698, 1983; U.S. Pat. No.5,985,579, which are incorporated herein by reference. A similardifferential screening modality can be used to generate binding agentsthat are aptamers, novel peptides, or artificial binding substrates.

The interaction of the bifunctional tag/target component complex with abinding agent can be monitored with a binding sensor using one or moreof the sensor methodologies disclosed herein. In this instance, thebinding agent configured to bind a bifunctional tag/target componentcomplex is attached to a component of the sensor. Upon binding of thebifunctional tag/target component complex, the sensor is activated andsends a signal to the controller indicating capture and sequestration ofthe target component. In response, the controller can optionallyinitiate release of one or more additional reactive components toinactivate and/or disrupt the sequestered target component.

Reactive Components for Inactivation of Target Components

The device disclosed herein further includes one or more secondreservoirs configured to release one or more reactive components formodulating a physiological effect of one or more sequestered targetcomponents. A reactive component includes, but is not limited to, adenaturing agent, a degradative agent, a cell-disrupting agent, amodulator, an energy source, an apoptotic agent, a programmed celldeath-inducing agent, a cytotoxic agent, cytostatic agent, achemotherapeutic agent, an antibody toxin or a combination thereof.Reactive components for modulating a physiological effect of one or moresequestered target components can be any of a number of chemical typesincluding but not limited to a a protein, a peptide, a small molecule, achemical, a toxin, an aptamer, or an inhibitory RNA, DNA, or othernucleic acid. The one or more reactive components are incorporated intoor released within one or more second reservoirs associated with thedevice. Alternatively, the one or more reactive components arediffusible components released from a reservoir of the device into theblood of the subject.

In an aspect, the reactive component can be a recombinant protein orpeptide. The recombinant protein or peptide can be generated exogenouslyand incorporated into one or more second reservoirs of the device. In anaspect, the recombinant protein or peptide can be generated by one ormore cells incorporated into the device. The one or more cells can begenetically modified to synthesize and secrete the one or more reactivecomponents. Cells that can be used for this purpose include, but are notlimited to, mammalian cells, enucleated cells (e.g., erythrocytes),plants cells, bacteria, or yeast. DNA sequences corresponding to one ormore reactive components are cloned into an appropriate cell type usingstandard procedures with appropriate expression vectors and transfectionprotocols. The genetically modified cells are encapsulated in one ormore compartments of the device and secrete the one or more reactivecomponents into the blood fluid or lymph fluid of a vertebrate subject.The genetically modified cells are kept separate from the circulation ofa vertebrate subject using a size-limiting biocompatible mesh ormembrane filter, for example, that allows passage of the cytotoxic,cytostatic, and/or apoptotic, but not the larger cells.

In an aspect, the one or more reactive components are released fromsynthetic vesicles or particles. Examples include any of a number ofdrug delivery vehicles including, but not limited to, phospholipidvesicles (liposomes), nanoparticles, polymers, or hydrogels. The releaseof the one or more reactive components can be triggered by binding of aspecific target to the synthetic vesicle or particle. For example, oneor more DNA aptamers can be incorporated into hydrogel and designed tobind one or more specific targets and release the contents of thehydrogel in response to the controller which releases the hydrogel intothe blood fluid or lymph fluid of the subject.

Reactive components can include denaturing agents that modulate aphysiological effect of the one or more target components. The deviceincluding one or more reactive components can include one or moredenaturing agents. The physiological effect of one or more targetcomponents can be modulated by the process of denaturation in which thesecondary, tertiary or quaternary structure of one or more targetcomponents are altered by denaturing agents. Examples of denaturingagents include, but are not limited to, acids such as acetic acid,trichloroacetic acid

(TCA), sulfosalicyclic acid, picric acid; solvents such as methanol,ethanol, and acetone; cross-linking agents such as formaldehyde andgluteraldehyde; chaotropic agents such as urea, guanidinium chloride,and lithium perchlorate; and disulfide bond reducers such as2-mercaptoethanol, dithithreitol, and TCEP. In an aspect, acids can beused to denature a protein molecule by exposing the protein molecule toa pH below its isoelectric point. Under these conditions, the proteinmolecule will lose its negative charge and retain only positive charges.The like positive charges can repel one another and in areas of largecharge density, the intramolecular repulsion can be sufficient enough tocause unfolding of the protein. The one or more denaturing agents can beincorporated into or released within one or more second reservoirs ofthe device. Alternatively, the one or more denaturing agents can bereleased by the device as diffusible agents into the blood.

Reactive components can include degradative agents that modulate aphysiological effect of the one or more target components. Thephysiological effect of one or more target components can be modulatedby the one or more degradative agents that act by breaking peptide bondswithin the primary amino acid sequence of the one or more inflammatorymediators. The one or more degradative agents can include any of anumber of agents designed to cleave one or more peptide bonds of theprimary amino acid sequence of one or more target components. Examplesof degradative agents, include but are not limited to proteases, strongacids, strong bases, free radicals, natural or synthetic proteasomes, orphotoactivatable agents. The one or more degradative agents can beincorporated into or released within one or more second reservoirs ofthe device. Alternatively, the one or more degradative agents can bereleased by the device as diffusible agents into the blood.

The device including one or more degradative agents can include one ormore proteases. Examples of proteases include, but are not limited to,serine proteases, e.g., as trypsin, chymotrypsin, elastase, dipeptidylpeptidase IV, and subtilisin; cysteine proteases, e.g., papain,cathepsins, caspases, calpains; aspartic acid proteases, e.g., pepsin,renin, and HIV-proteases; metalloproteases, e.g. carboxypeptidases,aminopeptidases, and matrix metalloproteases, e.g. MMP1 through MMP28.The one or more proteases can be free in solution. Alternatively, theone or more proteases can be bound to a substrate. In an aspect, trypsincan be bound to glass beads. See, e.g., Lee, et al., J. Dairy Sci.,58:473-476, 1974, which is incorporated herein by reference.Alternatively, trypsin and other proteases can be bound to an agarosematrix. Sources of immobilized proteases including trypsin and pepsinare available from commercial sources (Pierce Chemicals, Rockford, Ill.;Applied Biosystems, Foster City, Calif.).

The device including one or more degradative agents can include anatural or synthetic complex of proteases. In an aspect, the one or moretarget components can be subject to degradation using proteasomes. Aproteasome is a naturally occurring large protein complex that containsmultiple subunits. The complex includes several protease activities, forexample, chymotrypsin-like activity, trypsin-like activity, glutamicacid protease activity, and threonine protease activity. Proteasomecomplexes can be purified from fractionated cells usingultracentrifugation through a 10-40% glycerol gradient. See, e.g.,Pervan, et al., Mol. Cancer. Res. 3:381-390, 2005, which is incorporatedherein by reference. Proteasomes can be isolated using a commerciallyavailable isolation kit. (Proteasome Isolation Kit, Human 539176-1KIT,Calbiochem (EMD Chemicals, Inc.; Gibbstown, N.J.).

The device including one or more degradative agents can include an agentthat selectively targets one or more target component for degradation.In an aspect, the one or more target components can be covalently taggedwith ubiquitin for selective destruction by proteasomes. Ubiquitin is asmall and highly conserved protein. An isopeptide bond links theterminal carboxyl of ubiquitin to the ε-amino group of a lysine residueof a protein targeted for degradation. The joining of ubiquitin to thetargeted protein is ATP-dependent. Three enzymes are involved,designated E1, E2 and E3. Initially, the terminal carboxyl group ofubiquitin is joined in an ATP-dependent thioester bond to a cysteineresidue on ubiquitin-activating enzyme (E1). The ubiquitin is thentransferred to a sulfhydryl group on a ubiquitin-conjugating enzyme(E2). A ubiquitin-protein ligase (E3) then promotes transfer ofubiquitin from E2 to the ε-amino group of a lysine residue of a proteinrecognized by that E3, forming an isopeptide bond. There are distinctubiquitin ligases with differing substrate specificity. In addition,some proteins have specific sequences termed a “destruction box” that isa domain recognized by a corresponding ubiquitin ligase. In general, E1,E2, and E3 can be isolated from natural sources or generated usingstandard molecular biology techniques and used to ubiquinate proteins invitro. See, e.g., Chen, et al., EMBO Rep. 2:933-938, 2001, which isincorporated herein by reference. In an aspect, the E2 ligase can begenetically engineered in such a manner as to recognize a specificsubstrate. See, e.g., Colas, et al., PNAS 97:13720-13725, 2000, which isincorporated herein by reference. The device including the treatmentregion can further include one or more genetically engineered E2 ligaseenzymes capable of adding ubiquitin to and facilitating degradation ofthe one or more target components in the blood of the subject.

In a further aspect, the ubiquitin can be indirectly associated with theone or more target components. In an aspect, the ubiquitin can be linkedto an antibody or aptamer structural component of the bifunctional tag.Binding of the ubiquitin-labeled antibody or aptamer to one or moretarget components can mark the protein conjugate for degradation byproteasomes.

The device including one or more degradative agents can include a strongacid. Acid hydrolysis can result in degradation of the one or moretarget components. In this aspect, strong acids such as hydrochloricacid or sulfuric acid can be used to break the carbon-nitrogen peptidebond. Degradation of one or more target components by acid hydrolysiscan be optionally performed in combination with elevated temperature, anitrogen atmosphere and/or microwave energy.

The device including one or more degradative agents can include one ormore reactive oxygen species. Examples of reactive oxygen speciesinclude, but are not limited to, singlet molecular oxygen, superoxideion, hydrogen peroxide, hypochlorite ion, hydroxyl radical. Reactiveoxygen species can react directly with proteins, targeting peptide bondsor amino acid side chains, for example, reacting with the one or moretarget components bound by an affinity binding component to a surface ofthe treatment region. See, e.g., Davies, Biochem. Biophys. Res. Commun.305:761-770, 2003, which is incorporated herein by reference. A numberof the reactions mediated by reactive oxygen species lead tointroduction of carbonyl groups into the protein which in turn canresult in inactivation of the protein by cleavage of the peptide boundto yield lower-molecular weight products, cross-linking of proteins toyield higher-molecular weight products, or loss of catalytic function orstructural function by distorting secondary and tertiary structure, orcombination thereof. Reactive oxygen species can induce a amidation,diamide, glutamate oxidation and/or proline oxidation which can lead tocleavage of peptide bonds. Reactive oxygen species can be formed by theinteraction of biological molecules with components including, but notlimited to, ionizing radiation, as a byproduct of cellular respiration,and dedicated enzymes such as NADPH oxidase and myeloperoxidase.

In an aspect, the device including one or more degradative agents caninclude reactive oxygen species that are singlet oxygen species. Singletoxygen can cause damage to both the side-chains and backbone of aminoacids, peptides, and proteins. See, e.g., Davies, Biochem. Biophys. Res.Commun. 305:761-770, 2003, which is incorporated herein by reference.Singlet oxygen species can react with tryptophan, tyrosine, histidine,methionine and/or cysteine and cystine residues within a polypeptide andcan cause increased susceptibility to proteolytic enzymes, an increasedextent/susceptibility to unfolding, changes in conformation, an increasein hydrophobicity, and changes in binding of co-factor and metal ions.In particular, the interaction of tyrosine with singlet oxygen speciescan lead to fragmentation or cleavage of the polypeptide. See, e.g.,Davies, Biochem. Biophys. Res. Commun. 305:761-770, 2003, which isincorporated herein by reference.

The device including one or more degradative agents can include one ormore singlet oxygen species generated by a photosensitizer, a chemicalwhich upon exposure to a given wavelength of light emits singlet oxygenspecies. Examples of photosensitizers include, but are not limited to,porphyrin derivatives such as porfimer sodium, which is excited by redlight at 630 nm; chlorins and bacteriochlorins such as bonellin (maximumabsorbance 625 nm), mono-L-aspartyl chlorine e6 (max abs 654),m-tetrahydroxyphenyl chlorine (mTHPC, max abs 652 nm), and tinetiopurpurin (SnET2, maximum absorbance 660 nm); benzoporphyrinderivatives such as veteroporfin (also labeled BPD-MA, maximumabsorbance 690 nm), 5-aminolaevulinic acid (ALA, porphoryin precursor toPpIX (maximum absorbance 635 nm)); texaphyrins such as lutetiumtexaphyrin (Lu-Tex, maximum absorbance 732), Phthalocyanines andnaphthalocyanines (maximum absorbance 670-780 nm); and cationicphotosensitizers such as rhodamine 123 and methylene blue. See, e.g.,Prasad (2003) Introduction to Biophotonics, John Wiley & Sons, Inc.Hoboken, N.J. Tunable quantum dots (QDs), especially those absorbing inthe wavelength range of 600 to 800 nm, also emit singlet oxygen speciesin response to light provided by the device. The tunable quantum dotscan be useful as photosensitizers. See, e.g., Samia, et al., Photochem.Photobiol. 82:617-625, 2006, which is incorporated herein by reference.

Reactive components can include cell-disrupting agents that modulate aphysiological effect of the one or more target components. The deviceincluding one or more reactive components can include one or morecell-disrupting agents. Examples of cell-disrupting agents include, butare not limited to, alcohols and other organic solvents such asmethanol, ethanol, isopropanol, and acetone; cross-linking aldehydessuch as formaldehyde and gluteraldehyde; oxidizing agents such as sodiumhypochlorite, calcium hypochlorite, chloramine, chlorine dioxide,hydrogen peroxide, iodine, ozone, acidic electrolyzed water, peraceticacid, performic acid, potassium permanganate, potassiumperoxymonosulfate; acids such as acetic acid, trichloroacetic acid(TCA), sulfosalicyclic acid, picric acid; phenolics such as phenol,O-phenylphenol, chloroxylenol, hexachlorophene, thymol; chaotropicagents such as urea, guanidinium chloride, and lithium perchlorate; anddisulfide bond reducers such as 2-mercaptoethanol, dithiothreitol; andquaternary ammonium compounds. For example, organic solvents such asmethanol, ethanol or acetone can disrupt a cell by solubilizing thelipids in the plasma membrane and allowing the soluble contents of thecell to be released. In an aspect, the one or more cell-disruptingagents are incorporated into or released within one or more secondreservoirs of the device. In an aspect, the one or more cell-disruptingagents are released by the device as diffusible agents into the blood.

The physiological effect of one or more target components can bemodulated by the one or more cell-disrupting agents that act by breakingpeptide bonds within the primary amino acid sequence of proteins andpeptides associated with one or more target components. In an aspect,the device including one or more cell-disrupting agents can include oneor more proteases. Examples of proteases include, but are not limitedto, serine proteases, e.g., as trypsin, chymotrypsin, elastase,dipeptidyl peptidase IV, and subtilisin; cysteine proteases, e.g.,papain, cathepsins, caspases, calpains; aspartic acid proteases, e.g.,pepsin, renin, and HIV-proteases; metalloproteases, e.g.carboxypeptidases, aminopeptidases, and matrix metalloproteases, e.g.MMP1 through MMP28. In one aspect, the one or more proteases are free insolution. In an aspect, the one or more proteases are bound to asubstrate.

The device including one or more cell-disrupting agents can include oneor more free radical reactive oxygen species. Examples of reactiveoxygen species include, but are not limited to, singlet molecularoxygen, superoxide ion, hydrogen peroxide, hypochlorite ion, hydroxylradical. Reactive oxygen species can react directly with proteinsassociated with the one or more target components, targeting peptidebonds or amino acid side chains. In an aspect, the device including oneor more cell-disrupting agents can include reactive oxygen species thatare singlet oxygen species. Singlet oxygen can cause damage to both theside-chains and backbone of amino acids, peptides, and proteins. See,e.g., Davies, Biochem. Biophys. Res. Commun. 305: 761-770, 2003, whichis incorporated herein by reference. The cell-disrupting agents canfurther include one or more singlet oxygen species generated by aphotosensitizer. Examples of photosensitizers various classes ofphotosensitizers have been disclosed herein. A number of cell typesincluding cancer cells and bacterial pathogens are at least partiallyinactivated in response to treatment with photosensitizers such asphthalocyanines, phenothiazines, and porphyrins. See, e.g., Miller, etal., Toxicol. Appl. Pharmacol. 224:290-299, 2007; Jori, et al., LasersSurg. Med. 38:468-481, 2006; Keefe, et al., Lasers Surg. Med.31:289-293, 2002, each of which is incorporated herein by reference.

Modulators can modulate a physiological effect of the one or more targetcomponents. The device can include one or more reactive components thatare one or more modulators that either directly or indirectly modulatethe activity of one or more target components in the blood of avertebrate subject. The one or more modulators can be incorporated intoor released within one or more second reservoirs of the device.Alternatively, the one or more modulators can be released by the deviceas diffusible agents into the blood. A modulator can alter, modify,reduce or eliminate the activity of one or more target components bypreventing the binding of one or more target components to theirrespective cognates. Alternatively, a modulator can alter, modify,reduce or eliminate the activity of one or more target components byinhibiting the enzymatic activity, e.g., phosphorylation activity, ofthe one or more target components. Alternatively, the one or moremodulators can indirectly alter, modify or eliminate the activity of oneor more target components by attenuating the gene expression of one ormore target components. In an aspect, the one or more modulators canindirectly alter or eliminate the activity of one or more targetcomponents by increasing the expression of endogenous antagonists of theone or more target components.

In general, the one or more modulator can be a protein, a peptide, asmall molecule, an aptamer, or an inhibitory RNA, DNA, or nucleic acid.Modulators are contemplated that either directly or indirectlyantagonize the activity of one or more target components and/orattenuate expression of one or more target components.

In an aspect, the one or more modulator is designed to block theactivity or binding properties of one or more target components that isan inflammatory mediator. Examples of modulators of inflammatorymediator activity and binding include but are not limited to antibodies,e.g., infliximab, adalimumab, basiliximab efalizumab; soluble receptors,e.g., etanercept, abatacept, alefacept; corticosteroids, e.g.,hydrocortisone, cortisone, prednisone, prednisolone, methylprednisolone,meprednisone, triamcinolone, paramethsone, fluprednisolone,betametasone, and dexamethasone; nonsteroidal anti-inflammatory drugs(NSAIDS), e.g., selective cycloxygenase (COX) inhibitors exemplified bycelecoxib, etoricoxib, meloxicam, and valdecoxib and non-selective COXinhibitors exemplified by diclofenac, difluisal, etodolac, fenoprofen,fluripofen, ibuprofen, indomethacin, ketoprofen, ketorolac, nabumetone,naproxen, oxaprozin, piroxicam, sulindac, tenoxica, tiaprofen, tolmetin,azapropazone, and carprofen; and, e.g., methotrexate, azathioprine,pennicillamine, hydroxychloroquine, chloroquine, cyclophosphamide,cyclosporine, mycophenolate mofetil, gold, and sulfasalazine.

Reactive components can include cytotoxic, cytostatic, apoptotic, and/orchemotherapeutic agents that modulate a physiological effect of the oneor more cellular target components. The device including one or morereactive component can include one or more of a cytotoxic, a cytostatic,an apoptotic, and/or a chemotherapeutic agent. Reactive components thatare cytotoxic, cytostatic, apoptotic, and/or chemotherapeutic agents arecontemplated that either directly or indirectly inactivate or kill oneor more target components or target cells. Examples of cytotoxic,cytostatic, apoptotic, and/or chemotherapeutic agents include but arenot limited to vinca alkaloids (e.g., vinblastine, vincristine,vinflunine, vindesine, vinorelbine); taxanes (e.g., docetaxel,larotaxel, ortataxel, paclitaxel, tesetaxel); epothilones (e.g.,ixabepilone); dihydrofolate reductase inhiitors (e.g., aminopterin,methotrexate, pemetrexed); thymidylate synthase inhibitors (e.g.,raltitrexed); adenosine deaminase inhibitor (e.g., pentostatin);halogenated/ribonucleotide reductase inhibitors (e.g., cladribine,clofarabine, fludarabine); thiopurine (e.g., thioguanine,mercaptopurine); thymidylate synthase inhibitors (e.g., fluorouracil,capecitabine, tegafur, carmofur, floxuridine); DNA polymerase inhibitors(e.g., cytarabine); ribonucleotide reductase inhibitor (e.g.,gemcitabine, hydroxyurea); hypomethylating agent (e.g., azacitidine,decitabine); camptotheca (e.g., camptothecin, topotecan, irinotecan,rubitecan, belotecan); podophyllum (e.g., etoposie, teniposide);anthracyclines (e.g., aclarubicin, daunorubicin, doxorubicin,epirubicin, idarubicin, amrubicin, pirarubicin, valrubicin, zorubicin);anthracenediones (e.g., mitoxantrone, pixantrone); nitrogen mustards(e.g., mechlorethamine, cyclophosphamide, chlorambucil, bendamustine,uramustine, estramustine); nitrosureas (e.g., carmustine, lomustine,fotemustine, nimustine, ranimustine, streptozocin); aziridines (e.g.,carboquone, thioTEPA, triziquone, triethylenemelamine); platinum (e.g.,carboplatin, cisplatin, nedaplatin, oxaliplatin, triplatin,tetranitrate, satraplatin); hydrazines (e.g., procarbazine); triazenes(e.g., dacarbazine, temozolomide, altretamine, mitobronitol);streptomyces (actinomycin, bleomycin, mitomycin, plicamycin);aminolevulinic acid/methyl aminolevulinate; efaproxiral; porphyrinderivatives (porfimer sodium, talaporfin, temoporfin, verteporfin);farnesyltransferase inhibitors, cyclin-dependent kinase inhibitors,proteasome inhibitors, phosphodiesterase inhibitors, IMP dehydrogenaseinhibitors, lipooxygenase inhibitors, PARP inhibitors, endothelinreceptor antagonists (e.g., atrasentan); retinoid X receptor (e.g.,bexarotine); sex steroid (e.g., testolactone); amsacrine, trabectedin,alitretinoin, tretinoin, arsenic trioxide, celecoxib, demecolcine,elesclomol, elsamitrucin, etoglucid, lonidamine, lucanthone,mitoguazone, mitotane, oblimersen, temsirolimus, vorinostat. Thecytotoxic agent can further be a biological agent, e.g., a peptide, aprotein, an enzyme, a receptor and/or an antibody. Examples ofbiological agents currently used to treat cancer include, but are notlimited to, cytokines such as interferon-α, interferon-γ, andinterleukin-2, an enzyme such as asparaginase, and monoclonal antibodiessuch as alemtuzumab, bevacizumab, cetuximab, gemtuzumab, rituximab, andtrastuzumab.

The device including one or more reactive components that are cytotoxic,cytostatic, apoptotic, and/or chemotherapeutic agents can include one ormore of an antibacterial drug. Examples of antibacterial drugs include,but are not limited to, beta-lactam compounds (e.g., penicillin,methicillin, nafcillin, oxacillin, cloxacillin, dicloxacilin,ampicillin, ticarcillin, amoxicillin, carbenicillin, and piperacillin);cephalosporins and cephamycins (e.g., cefadroxil, cefazolin, cephalexin,cephalothin, cephapirin, cephradine, cefaclor, cefamandole, cefonicid,cefuroxime, cefprozil, loracarbef, ceforanide, cefoxitin, cefmetazole,cefotetan, cefoperazone, cefotaxime, ceftazidine, ceftizoxine,ceftriaxone, cefixime, cefpodoxime, proxetil, cefdinir, cefditoren,pivoxil, ceftibuten, moxalactam, and cefepime); other beta-lactam drugs(e.g., aztreonam, clavulanic acid, sulbactam, tazobactam, ertapenem,imipenem, and meropenem); other cell wall membrane active agents (e.g.,vancomycin, teicoplanin, daptomycin, fosfomycin, bacitracin, andcycloserine); tetracyclines (e.g., tetracycline, chlortetracycline,oxytetracycline, demeclocycline, methacycline, doxycycline, minocycline,and tigecycline); macrolides (e.g., erythromycin, clarithromycin,azithromycin, and telithromycin); aminoglycosides (e.g., streptomycin,neomycin, kanamycin, amikacin, gentamicin, tobramycin, sisomicin, andnetilmicin); sulfonamides (e.g., sulfacytine, sulfisoxazole,silfamethizole, sulfadiazine, sulfamethoxazole, sulfapyridine, andsulfadoxine); fluoroquinolones (e.g., ciprofloxacin, gatifloxacin,gemifloxacin, levofloxacin, lomefloxacin, moxifloxacin, norfloxacin, andofloxacin); antimycobacteria drugs (e.g., isoniazid, rifampin,rifabutin, rifapentine, pyrazinamide, ethambutol, ethionamide,capreomycin, clofazimine, and dapsone); and miscellaneous antimicrobials(e.g., colistimethate sodium, methenamine hippurate, methenaminemandelate, metronidazole, mupirocin, nitrofurantoin, polymyxin B,clindamycin, choramphenicol, quinupristin-dalfopristin, linezolid,spectrinomycin, trimethoprim, pyrimethamine, andtrimethoprim-sulfamethoxazole).

The device including one or more reactive components that are cytotoxic,cytostatic, apoptotic, and/or chemotherapeutic agents can include one ormore of an antifungal agent. Examples of antifungal agents include, butare not limited to, anidulafungin, amphotericin B, butaconazole,butenafine, caspofungin, clotrimazole, econazole, fluconazole,flucytosine griseofulvin, itraconazole, ketoconazole, miconazole,micafungin, naftifine, natamycin, nystatin, oxiconazole, sulconazole,terbinafine, terconazole, tioconazole, tolnaftate, and/or voriconazole.

The device including one or more reactive components that are cytotoxic,cytostatic, apoptotic, and/or chemotherapeutic agents can include one ormore of an anti-parasite agent. Examples of anti-parasite agentsinclude, but are not limited to, antimalaria drugs (e.g., chloroquine,amodiaquine, quinine, quinidine, mefloquine, primaquine,sulfadoxine-pyrimethamine, atovaquone-proguanil, chlorproguanil-dapsone,proguanil, doxycycline, halofantrine, lumefantrine, and artemisinins);treatments for amebiasis (e.g., metronidazole, iodoquinol, paromomycin,diloxanide furoate, pentamidine, sodium stibogluconate, emetine, anddehydroemetine); and other anti-parasite agents (e.g., pentamidine,nitazoxanide, suramin, melarsoprol, eflornithine, nifurtimox,clindamycin, albendazole, and tinidazole).

The device including one or more reactive components that are cytotoxic,cytostatic, apoptotic, and/or chemotherapeutic agents can include one ormore of an antiviral agent. Examples of antiviral agents include, butare not limited to, nucleoside analogs used to treat herpes simplexvirus (HSV) and varicella-zoster virus (VZV) (e.g., valacyclovir,famciclovir, penciclovir, and trifluridine); nucleoside analogs used totreat cytomegalovirus (CMV) (e.g., ganciclovir, valganciclovir, andcidofovir); nucleoside and nonnucleoside reverse transcriptaseinhibitors used to treat HIV (e.g., abacavir, didanosine, emtricitabine,lamivudine, stavudine, tenofovir, zalcitabine, zidovudine, delavirdine,efavirenz, and nevirapine); protease inhibitors used to treat HIV (e.g.,atazanavir, darunavir, fosamprenavir, indinavir, lopinavir, nelfinavir,ritonavir, saquinavir, and tipranavir); and drugs used to treathepatitis (e.g., interferon alfa, adefovir dipivoxil, entecavir, andribavirin).

The device can include one or more reservoirs that store one or morereactive components. The one or more reservoirs of the device areconfigured to controllably release one or more reactive components. Eachreservoir can contain one or more reactive components. Release of thereactive component from a reservoir is controlled by the controllercomponent of the device. In an aspect, the reactive components can behoused in multiple reservoirs associated with the device. For example,the device can include one or more microchips each with multiplereservoirs sealed with removable caps to enable controlled release ofone or more a denaturing agent, a degradative agent, a cell-disruptingagent, a modulator, an energy source, an apoptotic agent, a programmedcell death-inducing agent, a cytotoxic agent, cytostatic agent, achemotherapeutic agent, an antibody toxin or a combination thereof. See,e.g., U.S. Pat. No. 7,413,846; Maloney & Santini, Proceedings 26^(th)Annual International Conference IEEE EMBS, San Francisco, Calif., USA,Sep. 1-5, 2004, each of which is incorporated herein by reference.

Energy sources can modulate a physiological effect of the one or moretarget components. The device can include one or more reactivecomponents that include one or more energy sources configured tomodulate a physiological effect of the of one or more target components.The one or more energy sources can be directed to blood within thesecond reservoir or can be directed outside the device to the blood. Theone or more energy sources provide energy types including, but notlimited to, electromagnetic radiation, e.g., ultraviolet, infrared,optical, microwave, or millimeter wave; acoustic energy, e.g.,ultrasonic acoustic energy; heat; atmospheric pressure glow discharge;electron beam radiation; or gamma radiation. In an aspect, the energysource itself can modulate a physiological effect of one or more targetcomponents. Alternatively, heat generated by the energy source canmodulate a physiological effect of the one or more target components.

The application of one or more energy sources to the blood in the formof electromagnetic, acoustic, and/or electronic energy can inducedenaturation and/or degradation of one or more target components. Anenergy source can denature target component by unfolding the structureand/or inducing changes in amino acid chains and/or other side chains.The energy source can degrade a target component by cleaving one or morechemical bonds such as peptide bonds. The energy source can otherwisemodulate a physiological effect of the one or more target components byinducing aggregation of the one or more target components. The energysource can induce cellular disruption of a cellular target componentleading to inactivation, apoptosis or death of the target cell.

The device including the one or more energy sources can provide a set ofdiffering energy inputs specifically directed to modulating aphysiological effect of one or more target components. The set ofdiffering energy inputs selectively resonates a plurality of resonantstructures in the one or more target components and can modulate aphysiological effect of the one or more target components. See, e.g.,U.S. Patent Application 2007/0021927 A1, which is incorporated herein byreference. The differing energy inputs are selected to resonate one ormore resonant structures among the group of proximate atoms comprisingthe one or more target components. Application of a series of differingenergy inputs can have a physical effect, such as transferringsubstantially more energy to a group of proximate atoms relative toother atoms in the surrounding medium, breaking a predetermined bondbetween two members of the group of proximate atoms, or changing akinetic parameter of a reaction involving a member of the group ofproximate atoms. The one or more resonant structures can be resonatedsimultaneously, sequentially, and/or in a temporally overlappingfashion. The series of differing energy inputs can be appliedsimultaneously, sequentially, and/or in a temporally overlappingfashion.

The set of differing energy inputs can be electromagnetic beams, each ofwhich can have one or more characteristics including, but not limitedto, a selected set of frequencies, a selected set of phases, a selectedset of amplitudes, a selected temporal profile, a selected set ofpolarizations, or a selected direction. The temporal profile of the setof differing energy inputs can be characterized by a selected beamduration, and/or by a selected change in frequency, modulationfrequency, phase, amplitude, polarization, or direction during aselected time interval. At least one electromagnetic beam can bepolarized, amplitude modulated, or frequency modulated, and it can be,for example, an infrared beam. A plurality of electromagnetic beams candiffer in frequency, modulation frequency, phase, amplitude,polarization, or direction, and/or can intersect at a target location.The method can include scanning at least one electromagnetic beam.

In an aspect, the device can include reactive components that include anenergy field including, but not limited to, an electric field, amagnetic field, an electromagnetic field, a mechanical stress, amechanical strain, a lowered or elevated temperature, a lowered orelevated pressure, a phase change, an adsorbing surface, a catalyst, anenergy input, or a combination of any of these. The energy field canresult in inactivation of the one or more target components. Mechanicalstress, mechanical strain, lowered or elevated pressure, phase change,or adsorbing surface can provide energy to result in cellular disruptionof target cells.

The device including the one or more energy sources can generate heatthat induces inactivation of one or more target components. Exposure ofmost proteins or peptides to high temperature results in irreversibledenaturation due to the weakening of long range bonds associated withtertiary structure and cooperative hydrogen bonds associated withhelical structure. As these noncovalent bonds are broken, the proteinmolecule becomes more flexible and exposed to the solvent. Watermolecules associated with the solvent form new hydrogen bonds thatcannot be energetically overcome even as the protein molecule is cooled,leaving the protein molecule in an altered or denatured state. In anaspect, the one or more target components can be inactivated bytreatment with a heat source. The heat source can be electrical, heatingthe entirety of the one or more second reservoirs of the device.Alternatively, the heat source can heat a substrate of the device towhich one or more target components are attached. Alternatively, theheat source can be more focused such as that experienced from exposureto focused electromagnetic energy, e.g. from a laser. In an aspect, theone or more target components can bind to specific binding agentsassociated with one or more carbon nanotubes, the latter of which canemit heat in response to near infrared (NIR) radiation. See, e.g., Kam,et al., PNAS 102:11600-11605, 2005, which is incorporated herein byreference.

The device including the one or more energy sources can generatemicrowave energy for use in modulating a physiological effect of one ormore target components. An energy source that incorporates microwaveenergy can result in degradation of the one or more target components.Microwaves are electromagnetic radiation with wavelengths between 0.01and 1 meter and a frequency range between 0.3 and 30 GHz. The efficiencyof microwave denaturation and/or degradation can be enhanced byincluding an acid, a protease, a chemical or combination thereof. Theeffects of microwave energy on peptide bond integrity can provide morethat just rapid heating and suggests that some non-heat component ofmicrowaves facilitates breakdown of the peptide bond. See, e.g., Lill,et al., Mass. Spectrometry Rev. 26:657-671, 2007, which is incorporatedherein by reference.

The device including the one or more energy sources can generate focusedultrasound energy for use in modulating a physiological effect of one ormore target components. Sonication in the form of focused ultrasoundenergy can result in degradation of the one or more target components.In an aspect, high intensity focused ultrasound produces cavitationbubbles that when collapsed yield very high localized pressures and hightemperatures along with shear forces, jets and shock waves. The localincrease in temperature and pressure can effectively denature proteins.See, e.g., Lopez-Ferrer, et al., J. Proteome Res. 7:3860-3867, 2008,which is incorporated herein by reference.

The device including the one or more energy sources can generate plasmaby atmospheric dielectric-barrier discharge for use in modulating aphysiological effect of one or more target components. Plasma generatedby atmospheric dielectric-barrier discharge can result in degradation ofthe one or target components. See, e.g., Hou, et al., IEEE Transactionson Plasma Science 36:1633-1637, 2008, which is incorporated herein byreference. Plasma is an ionized gas in which a certain proportion of theelectrons are free rather than bound to an atom or molecule. The plasmacan be generated by a non-thermal discharge at atmospheric pressure byapplication of high voltages across small gaps. The atmosphericdielectric-barrier discharge can be scalable from one millimeter to onemeter (Walsh, et al., IEEE Transactions on Plasma Science 36:1314-1315,2008, which is incorporated herein by reference. A plasma jet and/or aplasma brush can be used to degrade one or more target components.

The device including the one or more energy sources can generate highenergy radiation for use in modulating a physiological effect of one ormore target components. High energy radiation can result in degradationof the one or more target components. Gamma radiation in the range ofabout 2.0 kGy to about 23.0 kGy is able to denature protein in a dosedependent manner as determined by size chromatography. See, e.g.,Vuckovic, et al., J. Serb. Chem. Soc. 70:1255-1262, 2005, which isincorporated herein by reference. Sources of gamma radiation that can beincluded in the device include but are not limited to cobalt-60,cesium-137, and technetium-99.

The device including the one or more energy sources can generateelectron beam radiation for use in modulating a physiological effect ofone or more target components. Electron beam radiation can result indegradation of the one or more target components. Electron beam energyof 92.9 kGy induces physical changes and loss of protein antigenicity(Katial, et al., J. Allergy Clin. Immunol. 110:215-219, 2002, which isincorporated herein by reference. A nanoscale electron beam generatorcan be devised from a network array structure of carbon nanotubes. See,e.g., U.S. Pat. No. 7,355,334, which is incorporated herein byreference.

The device including one or more energy source can emit electricalenergy in a focused area within the treatment region to inactivate oneor more cellular target components. For example, cancer cells insuspension can be at least partially ablated using electrical pulsessufficient to induce irreversible electroporation of the cells. See,e.g., Miller et al. Technol. Cancer Res. Treat. 4:699-705, 2005, whichis incorporated herein by reference. In an aspect, at least partialinactivation of one or more cellular target components can be achievedby exposure to 10-30, 0.3 millisecond pulses at 500 to 2500 V/cm.

The device including one or more energy source can emit electromagneticenergy sufficient to modulate a physiological effect of one or morecellular target components. The electromagnetic energy can range over athe spectrum of frequencies from gamma ray, x-ray, ultraviolet, visible,near infrared, infrared, microwave, to radiowaves.

The device including one or more energy source can emit ultravioletradiation to modulate a physiological effect of one or more cellulartarget components. A number of pathogens are inactivated or killed byultraviolet germicidal irradiation. Ultraviolet light ranges from UVA(400-315 nm; long wave or ‘blacklight’), UVB (315-280 nm, medium wave),and UVC (<280 nm, short wave or ‘germicidal’). The bacterium Escherichiacoli is partially or completely inactivated by exposure to a UVelectromagnetic energy source at wavelengths of 100-280 nm. Escherichiacoli as well as Salmonella enteritidis is also inactivated using pulsedbroad-spectrum electromagnetic energy with high UV content from, forexample, a Xenon lamp. In this instance, targeted bacteria are subjectedto 100-1000 pulses of broad-spectrum light with each pulse lasting, forexample, 85 ns and having, for example, a power output of 10 MW. See,e.g., Anderson et al. IEEE Transactions on Plasma Science 28:83-88,2000; Hancock et al. IEEE Transactions on Plasma Science 32:2026-2031,2004, each of which is incorporated herein by reference. Viruses andfungi (e.g., Aspergillus flavus and Aspergillus fumigatus) are alsoinactivated by ultraviolet irradiation. See, e.g., Tseng & Li, J. Occup.Envirn. Hyg. 4:400-405, 2007; Green et al. Can. J. Microbiol.50:221-224, 2004, each of which is incorporated herein by reference.

The device including one or more energy source can emit visible light tomodulate a physiological effect of one or more cellular targetcomponents. Staphylococcus aureus and Pseudomonas aeruginosa can beinactivated using a wavelength of 405 nm at doses ranging from 1-20J/cm². See, e.g., Guffey et al. Photomed. Laser Surg. 24:680-683, 2006,which is incorporated herein by reference. Pseudomonas aeruginosa aswell as Escherichia coli are partially inactivated using a wavelength of630 nm at 1-20 J/cm². See, e.g., Nussbaum et al. J. Clin. Laser Med.Surg. 20:325-333, 2002, which is incorporated herein by reference. In anaspect, a pathogen such as Escherichia coli can be at least partiallyinactivated or killed using a 810 nm diode laser with doses ranging from130-260 J/cm². See, e.g., Jawhara et al. Lasers Med. Sci. 21:153-159,2006, which is incorporated herein by reference. In some aspect, visibleor near infrared energy (e.g., 465 nm, 600 nm, and 950 nm) can be usedto alter the structural function of iron dependent pathogens by alteringthe function of heme iron prophyrins. See, e.g., U.S. Pat. No.6,030,653, which is incorporated herein by reference. In an aspect,viruses can be at least partially inactivated using a very low powerlaser emitting 80 femtosecond pulses at a wavelength of 425 nm andfrequency of 80 MHz. See, e.g., Tsen et al. Virol. J. 4:50, 2007, whichis incorporated herein by reference.

In an aspect, visible light energy can be combined with one or morereactive component that include a photosensitive agent to modulate aphysiological effect of one or more target cells. See, e.g., MaischLasers Med. Sci. 22:83-91, 2007; Jori et al. Lasers Surg. Med.38:468-481, 2006, each of which is incorporated herein by reference. Thevisible light energy combined with the one or more photosensitive agentscan be focused to a site of bacterial infection in the vertebratesubject, or can be focused onto the target cell or target componentwithin the target region in the vertebrate subject. For example,Staphylococcus aureus and Pseudomonas aeruginosa are inactivated usingeither a 0.95-mW helium-neon laser (632 nm) or a 5-mWindium-gallium-aluminum-phosphate laser (670 nm) with exposure dosesranging from 0.1 to 10.0 J/cm² in combination with the bacterialsensitizing agent, toluidine blue O. See e.g., DeSimone et al. Phys.Ther. 79:839-846, 1999, which is incorporated herein by reference.Similarly, bacterial inactivation by a laser diode or light-emittingdiode at 630 nm to 665 nm is enhanced in combination with methyleneblue. See e.g., Chan et al. Lasers Surg. Med. 18:51-55, 2003, which isincorporated herein by reference. A fluorescing dye, e.g., indocyaninegreen (ICG) can be used in combination with a diode laser emitting at808 nm to inactive a pathogen or pathogens. See e.g., Bartels et al.SPIE 2395:602-606, 1995, which is incorporated herein by reference. Inan aspect, a target cell, e.g., bacteria, can be inactivated using apolycationic photosensitizer in combination with irradiation with alaser diode or light-emitting diode at 630 nm to 665 nm at doses rangingup to a total fluence of 160 J/cm², e.g., in four 40 J/cm² aliquots,with imaging taking place after each aliquot of light. See e.g., Hamblinet al. Photochem. Photobiol. 75:51-57, 2002, which is incorporatedherein by reference. In an aspect, a target cell, e.g., Staphylococcusaureus, can be at least partially inactivated using energy from anargon-ion pumped dye laser (wavelength of 630 nm with total light doseup to 180 J/cm², fluence rate 250 mW/cm², wherein the total light dosecan be provided in one or more lower light energy aliquots) incombination with 5-aminolevulinic acid or porphyrin sodium. Effectivephotokilling of a target cell, e.g., Staphylococcus aureus orEscherichia coli, by endogenous porphyrins or exogenous porphyrins canbe achieved by application of light at 400-450 nm at approximately 50J/cm². With 600-700 nm light, a 10-fold higher light dose can provide asimilar result for S. aureus killing. With dye laser light at 632.8 nm,50 J/cm² can provide 3 orders of decrease in the viability of S. aureus.With white light, 75 J/cm² can provide 2-3 orders of decrease of S.aureus viability. See e.g., Karrer et al. Lasers Med. Sci. 14:54-61,1999; Nitzan et al. Lasers Med. Sci. 14:269-277, 1999, each of which isincorporated herein by reference.

The device including the one or more energy sources can generate heatthat alters the structural function of one or more cellular targetcomponents. In an aspect, the structural function of one or morecellular target components can be altered by laser-induced thermalenergy. Lasers are commonly used to treat cancers including but notlimited to basal cell carcinoma and the very early stages of cervical,penile, vaginal, vulvar, and non-small cell lung cancer. See e.g.,National Cancer Institute Laser in Cancer Treatment FactSheet, 2004,which is incorporated herein by reference. In an aspect, the device caninclude one or more laser-type component capable of emittingelectromagnetic energy sufficient to alter the structural function ofcirculating tumor cells or other target cells. Examples include but arenot limited to a carbon dioxide (CO₂) laser (10,600 nm, 0.1-0.2 mmpenetration depth), a Yttrium-Aluminium-Garnet (YAG) laser withNeodymium (Nd, 1064 nm or 1320 nm, 3-4 mm penetration depth), Erbium(Eb, 2940 nm, with <0.1 mm penetration depth), or Holmium (Ho, 2070 nm),diode laser (600-1600 nm), argon laser (488 nm and 514 nm, 1-1.5 mmpenetration depth), or an excimer laser (180-350 nm, cell/tissuedisintegration). As an example, melanoma and cervical cancer cells canbe ablated with a CO₂ laser using a power output ranging from 40 W to 80W. See, e.g., Gibson, et al. Br. J. Surg. 91:893-895, 2004; Bekassy etal. Lasers. Surg. Med. 20:461-466, 1997; Norberto et al. Surg. Endosc.19:1045-1048, 2005; Hansen et al. Minim. Invasive Ther. Allied Technol.15:4-8, 2006, each of which is incorporated herein by reference.Laser-induced thermal energy generated by a CO₂ or Nd:YAG laser can alsobe used to at least partially inactivate a pathogen. See, e.g. Bartelset al. SPIE 2395:602-606, 1995; Yeo et al. Pure Appl. Opt. 7:643-655,1998; U.S. Pat. No. 6,030,653; Gronqvist et al. Lasers Surg. Med.27:336-340, 2000, each of which is incorporated herein by reference.

The device including the one or more energy sources can emitelectromagnetic energy in the form of x-rays to alter the structuralfunction of one or more cellular target components. In an aspect, thedevice can contain a miniature X-ray emitter, such as that described inU.S. Patent Application 2004/0218724 A1, which is incorporated herein byreference. In an aspect, the device can contain radioisotopes, e.g.,cobalt 60, cesium 137, or europium 152, that emit strong gamma rays andcan be used to ablate cancerous cells. Optionally, the device cancontain other intrinsically radioactive isotope such as those that mightbe used for brachytherapy, including, for example, iodine 125, iodine131, strontium 89, phosphorous, palladium, or phosphate. In an aspect,the device can include an energy source that is an electron beam-drivenx-ray source. For example, breast cancer cells can be ablated using aminiature electron beam-driven x-ray source at doses of 5 to 20 Gy. See,e.g., Ross et al. Breast Cancer Res. 7:110-112, 2005, which isincorporated herein by reference. A nanoscale electron beam generatorcan be devised from a network array structure of carbon nanotubes. See,e.g., U.S. Pat. No. 7,355,334, which is incorporated herein byreference.

The device including the one or more energy sources can generateelectromagnetic energy in the form microwave or radiofrequency waves toalter the structural function of one or more cellular target components.The microwave range includes ultra-high frequency (UHF) (0.3-3 GHz),super high frequency (SHF) (3-30 GHz), and extremely high frequency(EHF) (30-300 GHz) signals. Microwave radiation at a frequency of 29.8GHz (Ka-band), for example, can be used to selectively kill bacteriawith minimal damage to healthy human cells See, e.g., Arndt et al.Microwave radiation—Therapeutic application for cure of subcutaneousbacterial infections. Space Life Sciences, NASA Research and TechnologyDevelopment. Biennial Research and Technology Report, which isincorporated herein by reference.

The device including the one or more energy sources can use focusedultrasound to generate heat to alter the structural function of one ormore cellular target components. Ultrasound causes tissue damage throughconversion of mechanical energy into heat and through cavitation.High-intensity focused ultrasound (HIFU) uses short exposures of focusedultrasound that rapidly increases cellular temperature above 80 degreescentigrade and is used for ablation, for example, of hepatocellularcarcinoma, prostate carcinoma, bladder and kidney cancers. See, e.g.,Kennedy et al. Br. J. Radiology 76:590-599, 2003, which is incorporatedherein by reference.

Optionally, the device can emit a laser-generated stress wave sufficientto disrupt a biological target. For example, stress waves sufficient todisrupt cell membranes can be generated with an ArF (193 nm) or a KrF(248 nm) eximer laser. Blood mononuclear cells and red blood cells aredamaged using, for example, 5 pulses of pressure ranging from 700 to1000 bar. See e.g., Lee et al. IEEE Journal of Selected Topics inQuantum Electronics 5:997-1003, 1999, which is incorporated herein byreference.

Two or more reactive components can be combined modulate a physiologicaleffect of one or more target components. The device can include two ormore reactive components that have been combined to inactivate ordisrupt one or more target components. The two or more combined reactivecomponents can be one or more binding agent combined with one or more adenaturing agent, a degradative agent, a cell-disrupting agent, amodulator, an energy source, an apoptotic agent, a programmed celldeath-inducing agent, a cytotoxic agent, cytostatic agent, achemotherapeutic agent, an antibody toxin or a combination thereof.

In an aspect, the two or more reactive components of the device can beincorporated into a single biomolecule. For example, the first reactivecomponent can be a binding agent, e.g., an antibody, that includes asecond reactive component that is a degradative activity. Certainantibodies are capable of cleaving the amide bond of peptide bonds. See,e.g., Janda, et al., Science 241:1188-1191, 1988; Lacroix-Desmazes, etal., J. Immunol. 177:1355-1365, 2006; Ponomarenko, et al., PNAS,103:281-286, 2006; and U.S. Pat. No. 6,387,674, each of which isincorporated herein by reference. Alternatively, the first reactivecomponent can be a binding agent, e.g., an antibody, and includes asecond reactive component that is a reactive oxygen species. The one ormore bifunctional tag/target component complex can bind to one or morebinding agents that are catalytic antibodies capable of generating thereactive oxygen species H₂O₂ in response to UV radiation. See, e.g.,Wentworth, et al., Science 293:1806-181811, 2001; Wentworth, Science296:2247-2248, 2002; Wentworth, et al., Proc. Natl. Acad. Sci. USA, 97:10930-10935, 2000, each of which is incorporated herein by reference.One or more antibodies or other binding agents can be generated for bothbinding and degradation of one or more bifunctional tag.

In an aspect, the first reactive component can be a binding agent, e.g.,an antibody that includes a second reactive component that is a cellulartoxin. For example, the first reactive component can be an antibody thatbinds a cellular target component and the second reactive component canbe a photosensitizer which is activated upon exposure to electromagneticenergy. See, e.g., Serebrovskaya, et al., Proc. Natl. Acad. Sci. USA.106: 9221-9225, 2009, which is incorporated herein by reference. Inanother example, the first reactive component can be an antibodydirected against a cancer cell or other cellular target component andthe second reactive component can be one or more of an auristatin,inhibitors of tubulin polymerization. See, e.g., Ma, et al., Clin.Cancer Res. 12: 2591-2596, 2006, which is incorporated herein byreference.

In an aspect, the two or more reactive components of the device can beincorporated into a single biomolecule and can include a first componentthat is a binding agent, e.g., an aptamer, and a second component thatis a degradative agent, e.g., a protease. For example, one or moreproteases can be conjugated or chemically linked to one or moreoligonucleotide-based aptamers. The oligonucleotide-based aptamers aredesigned to bind bifunctional tags/target component complexes. Uponbinding to the oligonucleotide-based aptamers, the one or morebifunctional tags/target component complexes are brought into proximityto the one or more proteases resulting in proteolytic degradation of theone or more bifunctional tags/target component complexes. Examples ofproteases have been provided herein and can be linked tooligonucleotide-based aptamers using any of a number of methods forconjugating a polypeptide to an oligonucleotide. In a further aspect, apolypeptide protease can be conjugated to an oligonucleotide-basedaptamer using a streptavidin-biotin bridge by introducing a biotinylatedoligonucleotide into the aptamer sequence and linking it to abiotinylated protease through a streptavidin bridge. Alternatively, thepolypeptide protease can be conjugated to the oligonucleotide-basedaptamer using a thiol-maleimide linkage in which a carbon with anattached thiol group is placed on the aptamer and reacts with amaleimide group added to the C terminus of the protease. See, e.g.,Nitin, et al., Nucleic Acids Res. 32:e58, 2004, which is incorporatedherein by reference. A number of modified nucleotides are commerciallyavailable for use in synthesizing oligonucleotide aptamers with aminesor other side chains for cross-linking (TriLink Biotechnologies, SanDiego, Calif.; Sigma Aldrich, St. Louis, Mo.).

In a further aspect, the first reactive component can be a binding agentlinked to a second reactive component encapsulated in a tunable vesicle.For example, the second reactive component, e.g., a denaturing and/ordegradative agent, can be encapsulated in a tunable hydrogel. Thebinding of one or more bifunctional tags/target component complexes tothe first reactive component, e.g., binding agent, releases thedenaturing and/or degradative agent from the hydrogel. In an aspect,target-responsive hydrogels can be generated in which the contents ofthe hydrogel are selectively released in response to binding a specifictarget. The hydrogel can incorporate one or more binding agents that areantibodies. The hydrogel can release its contents in response to anantibody-antigen interaction. See, e.g., Miyata, et al., PNAS103:1190-1193, 2006, which is incorporated herein by reference. In anaspect, the target-responsive hydrogel can incorporate one or morebinding agents that are oligonucleotide-based aptamers and release itscontents in response to an aptamer-ligand interaction. See Yang, et al.,J. Am. Chem. Soc. 130:6320-6321, 2008, which is incorporated herein byreference. In the latter case, two or more distinct aptamers configuredto partially overlap during hybridization can be copolymerized into apolyacrylamide hydrogel. At least one of the two or more aptamersfurther binds to a specific target, e.g., bifunctional tag/targetcomponent complex. When the bifunctional tags/target component complexesbinds to the aptamer, the number of nucleotide bases available forhybridization between the overlapping aptamers is reduced, causing themto separate. This separation allows the hydrogel to dissolute andrelease its contents. A target responsive hydrogel can be generatedwhich incorporates aptamers that specifically recognize one or moreinflammatory mediators. The hydrogel itself can be loaded with one ormore proteases or other reactive components that are configured tomodulate a physiological effect of inflammatory mediators. The contentsof the hydrogel are released upon binding of the one or moreinflammatory mediators to the aptamers associated with the hydrogel. Ina further aspect, hydrogels can be used for molecular imprinting. See,e.g., Byrne et al., “Molecular imprinting within hydrogels,” AdvancedDrug Delivery Reviews, 54: 149-161, 2002, which is incorporated hereinby reference.

Substrates for One or More Reactive Components

The one or more reactive components including binding agents, denaturingagents, degradative agents, modulators, or combinations there of can befree in solution within one or more second reservoirs of the device.Lower concentration of the reactive components can be used to actlocally at the site of the treatment region. Alternatively, the one ormore reactive components can be immobilized on a solid substrate withinthe one or more second reservoirs of the device. The solid substrate canbe a matrix, e.g., a bead or filter that is added to one or more secondreservoirs of the device. Examples of applicable solid substratesinclude, but are not limited to, beads, particles, membranes,semi-permeable membranes, capillary, or microarrays. The solid substratecan be comprised of an inorganic material, e.g., glass, alumina, silica,silicon, zirconia, graphite, magnetite, semiconductors, or combinationsthereof. Alternatively, the solid substrate can be comprised of anorganic material, e.g., polysaccharides including agarose, dextran,cellulose, chitosan, and polyacrylamide, polyacrylate, polystyrene,polyvinyl alcohol, or combinations thereof. Alternatively, the one ormore specific binding agents or one or more reactive components can beassociated with a solid substrate that are cells, e.g., mammalian cells,enucleated erythrocytes, bacteria, or viral particles or vesicles suchas liposomes or other micellular vesicles.

In an aspect, the one or more reactive components, either free insolution or bound to a solid substrate, can be contained near the one ormore second reservoirs including the treatment region of the deviceeither by size exclusion using a filter or mesh near the treatmentregion, containment within a hydrogel or polymer, or by physicalattachment to the treatment region of the device. In this aspect, one ormore bifunctional tag/target component complexes present in the bloodcan bind to the one or more reactive components and can be sequesteredfor inactivation as the blood passes through the device.

The one or more reactive components can be bound to the solid substrateeither directly or indirectly. For example, the one or more reactivecomponents can be coupled to the solid substrate by covalent chemicalbonds between particular functional groups on the specific binding agent(e.g., primary amines, sulfhydryls, carboxylic acids, hydroxyls, andaldehydes) and reactive groups on the solid substrate. A variety ofactivating compounds and schemes for directly bonding ligands to solidsubstrates are known. Some examples include, but are not limited to,cyanogen bromide, cyanuric chlorde, epichlorohydrin, divinyl sulphone,p-toluenesulphonyl chloride, 1,1′-carbonyldiimidazole, sodiummeta-periodate, 2-fluoro-1-methylpyridiniumtoluene-4-sulphonate,glycidoxypropyl-trimethoxysilane and 2,2,2-trifluoroethanesulphonylchloride. For example, cyanogen bromide in base reacts with hydroxyl(OH) groups on agarose solid substrate to form cyanate esters orimidocarbonates. These groups readily react with primary amines undermild conditions resulting in a covalent coupling of the ligand to theagarose solid substrate. Reactive imidocarbonates can also be formed oncarbon nanotubes, for example, through reactive carboxyl groupsgenerated by treatment of the nanotubes with oxidizing agents. See,e.g., Bianco, et al., in Nanomaterials for Medical Diagnosis andTherapy. pp. 85-142. Nanotechnologies for the Live Sciences Vol. 10Edited by Challa S. S. R. Kumar, WILEY-VCH Verlag GmbH & Co. KGaA,Weinheim, 2007, which is incorporated herein by reference.Functionalization of silicon chips with carboxyl groups can besubsequently used to immobilize proteins in the presence ofN-ethyl-N′-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) andN-hydroxysuccinimide ester (NHS). See, e.g., Hu, et al., Rapid Commun.Mass Spectrom. 21:1277-1281, 2007, which is incorporated herein byreference.

The one or more reactive components may or may not have linking orspacer groups bound to the C-terminus which when present can be used tobind the specific binding agent to the solid substrate indirectly. Whenpresent the linking group can be a polymer or a monomer. A linking groupcan be a chain of from 1-10 amino acids. Other examples of linkinggroups include, but are not limited to, polyethylene glycol,polypropylene glycol, polyesters, polypeptides, polyethers,polysaccharides, glycidoxyalkyl, alkoxyalkyl, alkyl, glycidoxypropyl,ethyl, propyl, phenyl and methacryl; and silicon containing linkinggroups such as diethyl(triethoxysilylpropyl)malonate;3-mercaptopropyltrimethoxysilane; 3-aminopropyltrimethoxysilane;N-[(3-trimethoxysilyl)propyl]ethylenediaminetriacetic acid;p-(chloromethyl)phenyltrimethoxysilane; vinyltriethoxysilane;3-bromopropyltriethoxysilane; and 3-glycidoxypropyltrimethoxysilane.

In general, any of a number of homobifunctional, heterofunctional,and/or photoreactive cross linking agents can be used to conjugate oneor more reactive components to an appropriately derivatized substrate.Examples of homobifunctional cross linkers include, but are not limitedto, primary amine/primary amine linkers such as BSOCES((bis(2-[succinimidooxy-carbonyloxy]ethyl) sulfone), DMS (dimethylsuberimidate), DMP (dimethyl pimelimidate), DMA (dimethyl adipimidate),DSS (disuccinimidyl suberate), DST (disuccinimidyl tartate), Sulfo DST(sulfodisuccinimidyl tartate), DSP (dithiobis(succinimidyl propionate),DTSSP (3,3′-dithiobis(succinimidyl propionate), EGS (ethylene glycolbis(succinimidyl succinate)) and sulfhydryl/sulfhydryl linkers such asDPDPB (1,4-di-(3′-[2′pyridyldithio]-propionamido) butane). Examples ofheterofunctional cross linkers include, but are not limited to, primaryamine/sulfhydryl linkers such as MBS(m-maleimidobenzoyl-N-hydroxysuccinimide ester), Sulfo MBS(m-maleimidobenzoyl-N-hydroxysulfosuccinimide), GMBS(N-gamma-maleimidobutyryl-oxysuccinimide ester), Sulfo GMBS(N-γ-maleimidobutyryloxysulfosuccinimide ester),EMCS(N-(epsilon-maleimidocaproyloxy) succinimide ester), SulfoEMCS(N-(epsilon-maleimidocaproyloxy) sulfo succinimide), SIAB(N-succinimidyl(4-iodoacetyl)aminobenzoate), SMCC (succinimidyl4-(N-maleimidomethyl)cyclohexane-1-carboxylate), SMPB (succinimidyl4-(rho-maleimidophenyl) butyrate), Sulfo SIAB(N-sulfosuccinimidyl(4-iodoacetyl)aminobenzoate), Sulfo SMCC(sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate),Sulfo SMPB (sulfosuccinimidyl 4-(rho-maleimidophenyl) butyrate), andMAL-PEG-NHS (maleimide PEG N-hydroxysuccinimide ester);sulfhydryl/hydroxyl linkers such as PMPI (N-rho-maleimidophenyl)isocyanate; sulfhydryl/carbohydrate linkers such asEMCH(N-(epsilon-maleimidocaproic acid) hydrazide); and amine/carboxyllinkers such as EDC (1-ethyl-3-(3-dimethylaminopropyl) carbodiimidehydrochloride).

In an aspect, the one or more reactive components can be linked to asolid substrate through non-covalent interactions. Examples ofnon-covalent interactions include, but are not limited to,protein-protein interactions such as those between avidin/streptavidinand biotin, protein A and immunoglobulins, protein G andimmunoglobulins, or secondary antibodies with primary antibodies. Forexample, the one or more reactive components can be modified with biotinusing standard methods and bound to a solid substrate derivatized withstreptavidin. Alternatively, one or more reactive components can bemodified with streptavidin and bound to a solid substrate derivatizedwith biotin. A single chain antibody can incorporate streptavidin aspart of a fusion protein to facilitate attachment of the antibody to thesolid substrate via a biotin-streptavidin linkage. See, e.g., Koo, etal. Appl. Environ. Microbiol. 64:2497-2502, 1999, which is incorporatedherein by reference. Solid substrates such as beads or other particulatesubstrates derivatized with protein A, protein G, streptavidin, avidin,biotin, secondary antibodies are available from commercial sources(from, e.g., Pierce-Thermo Scientific, Rockford, Ill., Sigma-Aldrich,St. Louis, Mo.). In an aspect, the one or more reactive components canbind to the solid substrate through a non-covalent interaction and befurther cross-linked to the solid substrate using a cross-linking agent.

In an aspect, the one or more reactive components can be associated witha solid substrate that are cells, e.g., mammalian cells, enucleatederythrocytes, bacteria, or viral particles, or vesicles such asliposomes or other micellular vesicles. Cells and vesicles can bemodified with one or more reactive components using many of the samemethods as provided herein. One or more reactive components can be boundto cells or vesicles using one or more homobifunctional orheterofunctional cross-linkers through primary amines and carboxylgroups. Alternatively, cells can be modified with one or more reactivecomponents using a biotin-streptavidin bridge. For example, one or morereactive components can be biotinylated and linked to a non-specificallybiotinylated cell surface through a streptavidin bridge. An antibody,aptamer, or receptor can be biotinylated using standard procedures. Thesurface membrane proteins of a cell can be biotinylated using an aminereactive biotinylation reagent such as, for example, EZ-LinkSulfo-NHS-SS-Biotin (sulfosuccinimidyl2-(biotinamido)-ethyl-1,3-dithiopropionate; Pierce-Thermo Scientific,Rockford, Ill., USA. See, e.g., Jaiswal, et al. Nature Biotech.21:47-51, 2003; U.S. Pat. No. 6,946,127, which is incorporated herein byreference.

In an aspect, the one or more reactive components can be associated withlipid or micellular vesicles. In an aspect, the one or more reactivecomponents can be antibodies attached to a liposome. Antibodies can beadded to liposomes using cross-linking agents and protein A. See, e.g.,Renneisen, et al., J. Bio. Chem., 265:16337-16342, 1990, which isincorporated herein by reference. The liposomes are formed from drylipid in the presence of an aqueous solution, e.g., a buffer ofappropriate pH followed by extrusion through a high pressure devicefitted with a polycarbonate filter with the desired pore size to formliposomes of a specific size range. The liposomes are modified withN-succinimidyl 3-(2-pyridyldithio) propionate-modified protein A. Theone or more antibodies are linked to the liposomes through selectivebinding to the protein A. Alternatively, thiolated antibodies can becovalently linked to liposomes prepared with 4-(p-maleimidophenyl)butyrylphosphatidyl-ethanolamine. See, e.g., Heath, et al., PNAS80:1377-1381, 1983, which is incorporated herein by reference.

In an aspect, the one or more reactive components can be expressed onthe surface of a cell. The one or more reactive components can benaturally expressed on the surface of a cell, such as a receptor of aspecific inflammatory mediator on a specific cell type. Alternatively,the one or more reactive components can be expressed on the surface of acell using genetic manipulation. For example, cells can be geneticallymanipulated to express a receptor that binds one or more structuralelements of the bifunctional tag. In one example, cells can begenetically manipulated to express one or more specific antibodies onthe cell surface. Methods have been provided for cell surface expressionof single chain Fv antibody fragments (scFv) fused tomembrane-associated proteins. See, e.g., Ho, et al., Proc. Natl. Acad.Sci. USA 103:9637-9642, 2006; Francisco, et al., Proc. Natl. Acad. Sci.USA 90:10444-10448, 1993; U.S. Pat. Appl. No. 2006/0083716, each ofwhich is incorporated herein by reference. In this aspect, the cDNAsequence encoding all or part of an antibody recognizing a structuralelement of a bifunctional tag is fused in an expression construct inframe with a membrane-associated protein and expressed in an appropriatecell type.

PROPHETIC EXAMPLES Example 1 Device Including Bifunctional Tags forSensing and Sequestering Inflammatory Mediators for Treatment of anInflammatory Condition or Disease

A device is described for treating an inflammatory condition orinflammatory disease associated with elevated levels of tumor necrosisfactor alpha (TNF-α) in the blood fluid or lymph fluid of a vertebratesubject. The device includes one or more aptamer-based piezoelectricsensors to sense target TNF-α in the blood fluid or lymph fluid of thevertebrate subject and to signal a controller on the device. Acontroller is configured to control flow of the blood fluid or lymphfluid through a lumen of the device in response to the aptamer-basedpiezoelectric sensor having sensed elevated levels target TNF-α in theblood fluid or lymph fluid. Upon detection of elevated levels of TNF-αin the blood fluid or lymph fluid, the controller responds by signalinga first set of reservoirs to release one or more bifunctional tags thatinclude antibodies directed against TNF-α, which are structural elementscapable of binding TNF-α. The device further includes binding agentsconfigured to bind the bifunctional tag/target component complex toremove it from the blood fluid or lymph fluid. The binding agents forcapturing the bifunctional tag/target component complex are within asecond set of reservoirs of the device. The device includes thecontroller in communication with the sensors, wherein the controller isconfigured to control flow of the blood fluid or lymph fluid through alumen of the device and to adjust release of the bifunctional tags tobind and sequester TNF-α to achieve a target value of TNF-α, that is, areduced level of TNF-α compared to elevated levels in the blood fluid orlymph fluid as a result of the inflammatory condition or inflammatorydisease in the vertebrate subject. The device includes a receiver forreceiving and processing data regarding the sensed levels of TNF-α and atransmitter for transmitting data to an external controller, a computingdevice, a physician, or a caregiver.

The device is placed in or proximal to one or more blood vessels of avertebrate subject. The device is a hollow stent-like structure that isplaced into a vessel at or near the site of inflammation using acatheter guide wire. The components of the device, including sensors,first and second reservoirs, controller and binding agents, are affixedto and/or incorporated into one or both surfaces of the stent-likestructure. The device is configured such that blood fluid or lymph fluidin the vessel is allowed to flow through the lumen either restricted bythe controllable flow barrier and the controller, or essentiallyunobstructed.

The device includes one or more piezoelectric sensors that sense thelevels of TNF-α in the blood of the vertebrate subject. Thepiezoelectric sensors respond to signals from aptamers astarget-recognition elements directed against TNF-α. Methods forgenerating aptamers against TNF-α using Systemic Evolution of Ligands byExponential Enrichment (SELEX) have been described. See, e.g., U.S. Pat.No. 7,309,789, which is incorporated herein by reference. Theinteraction of TNF-α with the aptamer target recognition elementstriggers the piezoelectric sensor to send a signal to the controller.The controller is an integral component of the device. The controllercalculates the levels of TNF-α in the blood based on the input from thesensors and compares these data with target values, e.g., desiredconcentrations of TNF-α. The level of TNF-α in a normal human subject isless than about 5 pg/ml. By contrast, a human subject experiencingseptic shock may have elevated levels of TNF-α of 140 pg/ml or higher.See, e.g., Ueda, et al., Am. J. Respir. Crit. Care Med. 160:132-136,1999 which is incorporated herein by reference. The target value forTNF-α may be that observed in a normal subject not experiencing aninflammatory disease or a disease resulting in an inflammatory response.In other instances, the target value for TNF-α may represent a reductionof at least 60%, relative to the current level of TNF-α in the blood ofthe subject. The controller sends a wireless signal to an externalcontroller to alert the subject and/or one or more caregivers as to thelevels of TNF-α in the blood of the subject.

The device including the controller releases one or more bifunctionaltags from the first reservoir of the device in response to sensingelevated levels of TNF-α in the blood fluid or lymph fluid of avertebrate subject. The bifunctional tags are stored and controllablyreleased from one or more micro-reservoirs incorporated into the device.The micro-reservoirs are covered by a seal that is disrupted in responseto an electrical signal from the controller. See, e.g., U.S. Pat. No.7,413,846; Maloney & Santini, Proceedings 26^(th) Annual InternationalConference IEEE EMBS, San Francisco, Calif., USA, Sep. 1-5, 2004, eachof which is incorporated herein by reference.

The bifunctional tags include a first structural element configured tobind TNF-α and a second structural element configured to interact with abinding agent of the device. The first structural element of thebifunctional tag is an antibody directed against TNF-α. Antibodies toTNF-α are available from commercial sources (from, e.g., Sigma-Aldrich,St. Louis, Mo.; or R&D Systems, Inc., Minneapolis, Minn.) or are readilygenerated using standard methods. The second structural element of thebifunctional tag is biotin configured to bind the binding agent,streptavidin. Biotin is incorporated into the TNF-α antibody andstreptavidin is bound to the treatment region of the device. Biotin isincorporated into the TNF-α antibody using standard methods andcommercially available labeling kits (e.g., NHS-PEO4-Biotinylation Kitfrom Thermo Fisher Scientific, Rockford, Ill.). Biotinylated TNF-αantibodies are also be available from commercial sources (e.g., fromBioLegend, San Diego, Calif.)

The device further includes one or more binding agents in the treatmentregion for capturing the bifunctional tag/target component complex. Thebinding agent is streptavidin configured to bind the biotin structuralelement of the anti-TNF-α antibody/biotin bifunctional tag in thetreatment region of the device. The binding agent, e.g., streptavidin,is incorporated into one or more second reservoirs of the device. Asecond reservoir of the device is part of the lumen through which all ora portion of the blood fluid or lymph fluid is diverted in response tothe sensor and controller sensing elevated levels of the targetcomponent. The streptavidin is coated on one or more surfaces of thesecond reservoir. Streptavidin is coated onto a metal surface using abiotinylated polyelectrolyte linker. See, e.g., Rossetti, et al.,European Cells Mater. 6 (Suppl. 1):83, 2003, which is incorporatedherein by reference. Alternatively, the streptavidin is coated onparticles, e.g., silica beads, magnetic beads, or polystyrene beads,which are incorporated into the second reservoir. Streptavidin coatedbeads are available from a variety of commercial sources (from, e.g.,Polysciences, Inc. Warrington, Pa.; Invitrogen, Carlsbad, Calif.; BangsLaboratories, Inc., Fishers, Ind.). As blood fluid or lymph fluid passthrough the lumen of the device, the biotinylated TNF-α antibody/TNF-αcomplexes bind to the streptavidin in the one or more second reservoirsand are removed from the peripheral circulation of the vertebratesubject. The antibody/TNF-α complexes are ablated in response toirradiation from a near-infrared energy source.

Example 2 Device Including Bifunctional Tags for Sensing andControllably Sequestering Inflammatory Mediators for Treatment of anInflammatory Condition or Disease

A device is described for treating an inflammatory condition orinflammatory disease associated with elevated levels of interleukin 6(IL-6) in the blood fluid or lymph fluid of a vertebrate subject. Thedevice includes a sensor that is an aptamer-based molecular beacon tosense IL-6 in the blood fluid or lymph fluid of the vertebrate subjectand to signal a controller on the device. A controller is configured tocontrol flow of the blood fluid or lymph fluid through a controllableflow barrier into a lumen of the device in response to the sensor havingsensed elevated levels of IL-6 in the blood fluid or lymph fluid. Theone or more bifunctional tags include one or more structural elementscapable of binding IL-6. The structural elements include antibodiesdirected against IL-6 and a binding element to the treatment region ofthe device. The device further includes binding agents configured tobind the bifunctional tag/target component complex to remove it from theblood fluid or lymph fluid. The binding agents for capturing thebifunctional tag/target component complex are within a second set ofreservoirs of the device. The binding agents are configured to only bindthe bifunctional tag when target component is bound. The device includesthe controller in communications with the sensors, wherein thecontroller is configured to control flow of the blood fluid or lymphfluid through a lumen of the device and to adjust release of thebifunctional tags to bind and sequester IL-6 to achieve a target valueof IL-6, that is, a reduced level of IL-6 compared to elevated levels inthe blood fluid or lymph fluid as a result of the inflammatory conditionor inflammatory disease in the vertebrate subject. The device includes areceiver for receiving and processing data regarding the sensed levelsof IL-6 and a transmitter for transmitting data to an externalcontroller, a computing device, a physician, or a caregiver.

The device includes one or more sensors configured to detect the levelsIL-6 in the blood of the vertebrate subject. The one or more sensors areaptamer-based molecular beacons designed to fluoresce in response toselectively binding one or more inflammatory mediators. Theaptamer-based molecular beacon includes a recognition element, afluorescing moiety, and a quenching moiety. The recognition elementselectively interacts with the target component, e.g., IL-6.Fluorescence induced by electromagnetic energy emitted by the device isquenched in the absence of IL-6. The binding of IL-6 to the selectiveaptamer induces a conformational change in the aptamer and increases thedistance between the fluorescing moiety and the quenching moietyresulting in a fluorescent signal in response to electromagnetic energy.The level of fluorescent signal is proportional to the level of IL-6 inthe blood fluid or lymph fluid. The emitted fluorescence is captured bya CCD or CMOS detector in the sensor and a corresponding signal is sentto the controller.

The one or more sensors is operably coupled to the controller, eitherwirelessly or by circuit, and can transmit data to the controllerregarding the detection and/or levels (relative or absolute) of IL-6 inthe blood fluid or lymph fluid of the subject. The controller is anintegral component of the device. The controller controls an energysource directed against the bound IL-6. The controller includes accessto stored data, or data that is stored off-site and coupled eitherwirelessly or by circuit to the sensor and the controller. Thecontroller also has access to one or more remote databases that includethe stored data. The stored data includes data regarding the normallevel of IL-6 in normal or healthy subjects without an inflammatorydisease or condition. The stored data includes data regarding thebaseline level of IL-6 in a subject prior to onset of the inflammatorydisease or condition. The stored data also includes data regarding thelevel of IL-6 in a subject at one or more previous time points. Thecontroller calculates the levels of IL-6 in the peripheral blood basedon the input from the sensors and compares these data with targetvalues, e.g., desired concentrations of IL-6. The target value for IL-6is that observed in a normal subject not experiencing an inflammatorydisease or a disease resulting in an inflammatory response. The targetvalue of IL-6 is a low serum level of IL-6 (mean of 15 pg/ml) ascompared to a high serum level of IL-6 (mean of 735 pg/ml), the latterof which may be associated with development of severe sepsis andincreased risk of death. See, e.g., Kellum, et al., Arch. Intern. Med.167:1655-1663, 2007; Presterl, et al., Am. J. Respir. Crit. Care Med.156:825-832, 1997; which are incorporated herein by reference. Thetarget value for IL-6 represents a reduction of at least 60% relative tothe current level of IL-6 in the peripheral blood of the subject. Thecontroller sends a wireless signal to an external controller to alertthe subject and/or one or more caregivers as to the levels of IL-6 inthe peripheral blood of the subject.

The controller releases the bifunctional tags from the first reservoirof the device in response to molecular beacon sensing elevated levels ofIL-6 in the blood fluid or lymph fluid of a vertebrate subject. Thebifunctional tags include a first structural element configured to bindIL-6 and a second structural element configured to interact with abinding agent of the device. The first structural element of thebifunctional tag is an antibody directed against IL-6. Antibodies toIL-6 are available from commercial sources (from, e.g., Sigma-Aldrich,St. Louis, Mo.; or R&D system, Inc., Minneapolis, Minn.) or are readilygenerated using standard methods. The second element of the bifunctionaltag is a conformational epitope generated by the binding of IL-6 to theanti-IL-6 antibody and selectively recognized by one or more bindingagents at a treatment region of the device.

The device further includes a conformation-specific antibody thatincludes binding agent for capturing the bifunctional tag/targetcomponent complex. The bifunctional tag is configured such that theconformation-specific antibody has an increased affinity for thebifunctional tag bound to the target component (e.g., antibody/IL-6complex) compared to an affinity for the bifunctional tag unbound to thetarget component. The conformation-specific antibody is configured torecognize a conformational epitope formed by the binding of IL-6 to ananti-IL-6 antibody. Conformation specific antibodies are generated thatselectively bind to an antibody-antigen complex but are unable to bindto either the antibody alone or the antigen alone. See, e.g., Nemazee &Sato, Proc. Natl. Acad. Sci, USA. 79:3828-3832, 1982; U.S. Pat. No.5,985,579, which are incorporated herein by reference. The bindingagent, e.g., the conformation specific antibody, is incorporated intoone or more second reservoirs of the device. The second reservoirincludes all or part of a lumen through which all or a portion of theblood fluid or lymph fluid has been diverted in response to sensingelevated levels of the IL-6 target component. The conformation-specificantibody is coated on one or more surfaces of the second reservoir andbinds the IL-6/anti-IL-6 antibody bifunctional reagent as it passesthrough the lumen of the device thus removing IL-6 from the peripheralcirculation of the vertebrate subject. The antibody/IL-6 complexes areablated in response to irradiation from a near-infrared energy source.

Example 3 Device Including Bifunctional Tags for Sensing, Binding andAltering Metastatic Tumor Cells for Treatment of a Neoplastic Disease orCondition

A device is described for treating a neoplastic disease or conditionincluding one or more light-scattering sensors to sense one or morecirculating tumor cells in blood fluid or lymph fluid of a vertebratesubject, a controller configured to release one or more bifunctionaltags from a first set of reservoirs in response to the light-scatteringsensor. The bifunctional tags are structural elements configured to bindcirculating lung tumor cells. The structural elements are antibodiesconfigured to selectively bind an epithelial cell-specific antigen, theepithelial cell-cell adhesion molecule (EpCAM), which is a tumor antigenspecific to metastatic tumor cells of epithelial origin. The controlleris further configured to control flow of the blood fluid or lymph fluidthrough a controllable flow barrier into a lumen of the device inresponse to the sensor, and a reactive component in a treatment regionincluding localized high concentrations of one or more cytotoxicchemotherapeutic agents configured to bind and inactivate the one ormore circulating tumor cells. The device is placed in or proximal to oneor more blood vessels and includes multiple lumens configured to receivea portion of the blood through a flow route, wherein the controllerconfigured to control flow of blood through the flow route into thelumen. The lumen includes one or more second reservoirs configured withconformational-specific antibodies to the anti-EpCAM/circulating lungtumor cell complex and are used to sequester the metastatic lung tumorcells within the second reservoir. The second reservoir further includesone or more reactive components including localized high concentrationsof one or more cytotoxic chemotherapeutic agents to induce apoptosis ornecrosis of the circulating metastatic tumor cells. The secondreservoirs can release the components of an AT chemotherapy regimen:doxorubicin, either alone or in combination with paclitaxel. The deviceincludes a receiver for receiving and processing data regarding thesensed levels of circulating metastatic lung tumor cells and atransmitter for transmitting data to an external controller, a computingdevice, a physician, or a caregiver.

The device includes one or more light-scattering sensors that sense thelevels of circulating metastatic lung tumor cells in the blood of avertebrate subject. Differential light scattering is used to detectcirculating tumor cells derived from metastasis of solid tumors. Ingeneral, a circulating metastatic lung tumor cell is characterized byits large size, immature appearance, increased nuclear to cytoplasmicratio, abnormally shaped nuclei, and disproportionately large nucleolusor multiple nucleoli. The size differential between a circulating tumorcell and components of the blood is used to specifically detect thecancerous cells. For example, the average diameter of neutrophils, redbloods cells, and platelets is 10.5-12.5 microns, 7-8 microns, and 3microns, respectively. In contrast, the average size of circulatingtumor cells isolated from subjects with breast, colon, stomach, and lungcancers range from 18.3 to 20.6 microns in diameter. Circulatingneuroblastoma tumor cells, for example, are greater than 20 microns indiameter. See, e.g., Moore et al. Cancer 13:111-117, 1960; Mohamed etal. IEEE Transactions on Nanobioscience, 3:251-256, 2004, each of whichis incorporated herein by reference. The size of a cell or cells passingby the one or more sensors is determined using forward and side lightscattering. The size, as measured in diameter, is compared with knownparameters regarding the size of normal blood components.

The controller calculates the number of circulating metastatic lungtumor cells in the blood fluid or lymph fluid based on the input fromthe sensors and compares these data with target values, e.g., reducedconcentrations of circulating metastatic lung tumor cells to a targetvalue of zero metastatic lung tumor cells in the blood fluid or lymphfluid. However, in some instances, simply lowering the number ofcirculating metastatic lung tumor cells may improve prognosis. Forexample, breast cancer patients with levels of circulating tumor cellsequal to or higher than five cells per 7.5 milliliters of blood have ashorter median progression-free survival (2.7 months vs. 7.0 months) andshorter overall survival (10 .1 months versus greater than 18.0 months)as compared with breast cancer patients with less than five cells per7.5 milliliters of blood. See, Cristofanilli et al. N. Engl. J. Med.351:781-791, 2004, which is incorporated herein by reference. As such,the target value for circulating tumor cells may represent a reductionof at least 60%, relative to the current level of circulating metastaticlung tumor cells in the blood of the subject.

The controller releases bifunctional tags, anti-EpCAM antibody complexedto a secondary binding agent, biotin, from one or more first reservoirsof the device in response to sensing elevated levels of circulatingtumor cells in the blood fluid or lymph fluid of a vertebrate subject.The bifunctional tag includes a first structural element anti-EpCAMantibody configured to bind a circulating tumor cell and a secondstructural element, biotin, configured to interact with a binding agent,streptavidin, in the reaction region of the device. The first structuralelement of the bifunctional tag is an antibody directed againstepithelial cell-cell adhesion molecule (EpCAM), an example of anepithelial cell-specific antigen commonly detected on metastatic tumorcells of epithelial origin. Antibodies to EpCAM are available fromcommercial sources (from, e.g., Sigma-Aldrich, St. Louis, Mo.; or R&DSystems, Inc., Minneapolis, Minn.) or are readily generated usingstandard methods. The second structural element of the bifunctional tagis biotin configured to bind a binding agent, streptavidin, of thedevice. All or part of the anti-EpCAM antibody and all or part of thebiotin may be crossed linked together to form a single bifunctional tag.The antibody can be cross-linked with the biotin to form a bifunctionaltag using a chemical cross-linking agent, e.g., N-succinimidyl3-(2-pyridyldithio)propionate (SPDP). See, e.g., U.S. Pat. No.5,470,570, which is incorporated herein by reference.

The device further includes one or more binding agents for capturing thebifunctional tag/circulating tumor cell complex. In this example, thebinding agent is streptavidin that binds the biotin structural elementof the bifunctional tag. The binding agent, e.g., streptavidin, isincorporated into the second reservoir of the device. The secondreservoir of the device may be all or part of a lumen through which atleast a portion of the blood fluid or lymph fluid has been diverted inresponse to sensing elevated levels of circulating metastatic lung tumorcells. In this example, the streptavidin is coated on one or moresurfaces of the second reservoir. For example, a silicon chip-likesurface can be functionalized with carboxyl groups and subsequently usedto immobilize proteins, e.g., streptavidin, in the presence ofN-ethyl-N′-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) andN-hydroxysuccinimide ester (NHS). See, e.g., Hu, et al., Rapid Commun.Mass Spectrom. 21: 1277-1281, 2007, which is incorporated herein byreference. Alternatively, the streptavidin can be used to coatparticles, e.g., silica beads, magnetic beads, or polystyrene beads, andincorporated into the second reservoir. Activated beads forcross-linking to proteins, e.g., streptavidin, are available from avariety of commercial sources (from, e.g., Polysciences, Inc.Warrington, Pa.; Invitrogen, Carlsbad, Calif.; Bangs Laboratories, Inc.,Fishers, Ind.). In this instance, the particles coated with thestreptavidin are retained in the treatment region due to either sizeexclusion or magnetic properties.

The controller controls flow of the blood fluid or lymph fluid into theone or more second reservoirs of the device. The bifunctionaltag/circulating tumor cell complexes present in the blood fluid or lymphfluid bind to the one or more binding agents in the second reservoir.Once sequestered in the second reservoir, the circulating tumor cellscan be further subjected to one or more reactive components to inducecell-disruption, apoptosis, and/or death of the tumor cells. The one ormore second reservoirs is configured to release localized highconcentrations of one or more cytotoxic agent, e.g., thechemotherapeutic agent doxorubicin (Adriamycin®), either alone or incombination with paclitaxel (Taxol®) resulting in apoptosis or necrosisof the lung tumor cells. The controller in the device maintains bloodflow through the treatment region of the device until reduced targetlevels of circulating lung tumor cells are reached in the blood fluid orlymph fluid of the subject.

Example 4 Device Including Bifunctional Tags for Sensing, Binding andAltering Inflammatory Mediators for Treatment of an InflammatoryCondition or Disease

A device is described for treating an inflammatory condition orinflammatory disease associated with elevated levels of eosinophils inthe blood fluid or lymph fluid of a vertebrate subject. The deviceincludes one or more piezoelectric sensor to sense eosinophils in bloodfluid or lymph fluid of a vertebrate subject and to signal a controlleron the device. The controller is configured to control flow of the bloodfluid or lymph fluid through a controllable flow barrier into multiplelumens of the device in response to the piezoelectric sensor havingsensed elevated levels of eosinophils in the blood fluid or lymph fluid.The bifunctional tags include one or more structural elements capable ofbinding eosinophils that include one or more ligands configured toselectively bind receptors on the surface of eosinophils. The one ormore structural elements include interleukin 5 (IL-5) directed againstthe eosinophil cell surface IL-5 receptor. The device is placed in orproximal to one or more vessels and includes multiple lumens configuredto receive at least a portion of the blood fluid or lymph fluid througha controllable flow barrier into a flow route, the controller configuredto control flow of blood fluid or lymph fluid through the flow routeinto the multiple lumens. The lumens include one or more secondreservoirs configured with one or more binding agents, e.g.streptavidin, for capturing and sequestering the bifunctionaltag/eosinophil complex. The second reservoir further includes one ormore reactive components, e.g., an energy source emitting near-infraredradiation, to induce thermal ablation of the sequestered eosinophils.The device includes a receiver for receiving and processing dataregarding the sensed levels of circulating eosinophils and a transmitterfor transmitting data to an external controller, a computing device, aphysician, or a caregiver. The device includes one or more sensors thatsense the levels of the eosinophils in the blood of the vertebratesubject. The sensors are piezoelectric sensors in which aptamers thatrecognize surface components of the eosinophils are used astarget-recognition elements. The interaction of eosinophils with theaptamer target-recognition elements triggers the piezoelectric sensor tosend a signal to the controller. The controller is an integral componentof the device. The one or more sensors is operably coupled to thecontroller, either wirelessly or by circuit, and can transmit data tothe controller regarding the detection and/or levels (relative orabsolute) of eosinophils in the blood fluid or lymph fluid of thesubject. The controller is an integral component of the device. Thecontroller controls other components of the device. The controllercontrols flow of the blood fluid or lymph fluid through a controllableflow barrier into multiple lumens of the device in response to thepiezoelectric sensor having sensed elevated levels of eosinophils in theblood fluid or lymph fluid. The controller controls an energy sourcedirected against the bound eosinophils. The controller includes accessto stored data, or data that is stored off-site and coupled eitherwirelessly or by circuit to the sensor and the controller. Thecontroller also has access to one or more remote databases that includethe stored data. The stored data includes data regarding the normallevel of eosinophils in normal or healthy subjects without aninflammatory disease or condition. The stored data includes dataregarding the baseline level of eosinophils in a subject prior to onsetof the inflammatory disease or condition. The stored data also includesdata regarding the level of eosinophils in a subject at one or moreprevious time points. The controller calculates the levels ofeosinophils in the blood based on the input from the sensors andcompares these data with target values, e.g., desired concentrations ofeosinophils. For example, the number of eosinophils in a normal humansubject ranges from about 45 cells/microliter to about 450cells/microliter. By contrast, a human subject experiencinghypereosinophilic syndrome (HES) has more than 1500eosinophils/microliter. See, e.g., Roufosse. Haematologica 94:1188-1193,2009, which is incorporated herein by reference. In some instances, thetarget value for eosinophils is that observed in a normal subject notexperiencing an inflammatory disease or a disease resulting in aninflammatory response. In other instances, the target value foreosinophils may represent a reduction of at least 60% relative to thecurrent level of eosinophils in the blood of the subject. The controllermay optionally send a wireless signal to an external controller to alertthe subject and/or one or more caregivers as to the levels ofeosinophils in the blood of the subject.

The controller controls release of bifunctional tags from the firstreservoir of the device in response to sensing elevated levels ofeosinophils in the blood fluid or lymph fluid of the vertebrate subject.The bifunctional tag includes a first structural element configured tobind eosinophils and a second structural element configured to interactwith a binding agent in a treatment region of the device. The firststructural element of the bifunctional tag is IL-5, the ligand for theIL-5 receptor expressed on the surface of eosinophils. The secondstructural element of the bifunctional tag is biotin configured to binda binding agent, streptavidin, of the device. A bifunctional tagincluding IL-5 and biotin is generated as a single fusion protein.Fusion proteins containing interleukins and other ligands can begenerated using standard protein engineering techniques. See, e.g., Hu,et. al., Canc. Res. 56:4998-5004, 1996, which is incorporated herein byreference.

The device further includes the binding agent, streptavidin, forcapturing the bifunctional tag/eosinophil complex. The streptavidin isconfigured to bind biotin, the second ligand structural element of thebifunctional tag. Streptavidin is incorporated into one or more secondreservoirs of the device. The second reservoir of the device is part ofa lumen through which at least a portion of the blood fluid or lymphfluid has been diverted in response to sensing elevated levels ofeosinophils. The streptavidin is coated onto an array of carbonnanotubes incorporated into a second reservoir of the device. Thestreptavidin is attached to a carbon nanotube by first functionalizingraw single-walled carbon nanotubes with amino groups using azomethineylide cycloaddition and then cross-linking a thiolated antibody to thefunctionalized nanotubes using a heterobifunctional cross-linker such assucciminidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxy-(6-amidocaproate).See, e.g., McDevitt, et al., J. Nucl. Med. 48:1180-1189, 2007, which isincorporated herein by reference.

The controller controls flow of the blood fluid or lymph fluid throughthe controllable flow barrier into the one or more second reservoirs ofthe device. The bifunctional tag/biotin/eosinophil complexes in theblood fluid or lymph fluid bind to the streptavidin on the carbonnanotube array in the second reservoir. Once sequestered in the secondreservoir, the bifunctional tag/biotin/eosinophil complex bound to thestreptavidin /carbon nanotube is subjected to one or more reactivecomponents, e.g., near infrared radiation, to induce cell-disruption,apoptosis, and/or death of the cells. The controller controls an energysource directed against the bound eosinophils. The reactive component,near infrared radiation, is used to induce carbon nanotube-mediatedthermal ablation of the sequestered eosinophils. Carbon nanotubes arecapable of converting near-infrared light into heat and can be used forhyperthermia ablation of targeted cells. See, e.g., Chakravarty, et al.,Proc. Natl. Acad. Sci. USA. 105:8697-8702, 2008, which is incorporatedherein by reference. As blood fluid or lymph fluid pass through thelumen of the device, the bifunctional tag/biotin/eosinophil complexesbind to the streptavidin/carbon nanotubes in the one or more secondreservoirs and are ablated in response to irradiation from anear-infrared energy source.

Example 5 Device Including Bifunctional Tags for Binding andSequestering an Illicit Drug for Treatment of Drug Addiction

A device is described for treating an overdose of illicit drugassociated with elevated levels of cocaine in the blood fluid or lymphfluid of a vertebrate subject. The device includes one or more sensorsto sense fluorescence associated with binding of an aptamer-basedmolecular beacon to a target component, e.g., cocaine, in the bloodfluid or lymph fluid of the vertebrate subject and a controllerconfigured to control flow of the blood fluid or lymph fluid through alumen in response to the sensor sensing elevated levels of cocaine inthe blood fluid or lymph fluid. The bifunctional tag includes one ormore structural elements capable of binding cocaine that include anaptamer-based fluorescent molecular beacon that responds to binding ofcocaine to the bifunctional tag. The structural element capable ofbinding cocaine is an aptamer-based fluorescent molecular beacondirected against cocaine. The device includes a sensor for sensingfluorescence associated with binding of cocaine to the aptamer-basedmolecular beacon. The sensor is in communication with the controllerthat is further configured to control flow of the blood fluid or lymphfluid through a controllable flow barrier into multiple lumens of thedevice in response to the sensor having sensed the bifunctionaltag/cocaine complex in the blood fluid or lymph fluid. The devicefurther includes binding agents in the treatment region of the device tobind the bifunctional tag/cocaine complex and to remove the complex fromthe blood fluid or lymph fluid. The device optionally includes areceiver for receiving and processing data regarding the sensed levelsof cocaine and a transmitter for transmitting data to an externalcontroller, a computing device, a physician, a corrections officer, or acaregiver.

The controller releases one or more bifunctional tags from one or morefirst reservoirs of the device in response to sensing cocaine in theblood of a vertebrate subject. The bifunctional tag includes the firststructural element, aptamer-based fluorescent molecular beacon directedagainst cocaine and a second structural element configured to interactwith a binding agent of the device. The second structural element is apeptide antigen configured to bind to an antibody binding agent in thetreatment region of the device. The cocaine-binding aptamer is furtherconfigured to serve as a molecular beacon, generating a fluorescentsignal upon binding cocaine. A cocaine-binding aptamer is generatedusing SELEX in which cocaine is immobilized on a column matrix andscreened through iterative rounds against a diverse library of DNAoligonucleotide sequences to find aptamers that selective bind cocaine.The aptamer is further modified by a fluorophore and a quencher togenerate a molecular beacon. The cocaine-binding aptamer can be modifiedwith fluorescein and a quencher such as, for example,4-[4′-((dimethylamino)phenyl)azo-] benzoic acid (DABCYL). See, e.g.,Strojanovic, et al. J. Am. Chem. Soc. 123:4928-4931, 2001, which isincorporated herein by reference. The first structural element, e.g.,cocaine-binding aptamer, and the second structural element, e.g.,peptide antigen, are covalently linked to form the bifunctional tag. Theaptamer-based molecular beacon is linked to a peptide using astreptavidin-biotin bridge, a thiol-maleimide linkage or disulfide bondsas described by Nitin, et al., Nucleic Acids Res. 32:e58, 2004, which isincorporated herein by reference.

The device includes one or more aptamer-based molecular beacon sensorsfor sensing the interaction of the bifunctional tag with the targetcomponent, e.g., cocaine. The sensors are configured to sensefluorescence emitted from bifunctional tag when the target component,e.g., cocaine, has bound to the aptamer-based molecular beacon. Thedevice provides an electromagnetic energy source for illuminating theblood or lymph vessel and a charged couple device (CCD) image capturecomponent for sensing emitted fluorescence. The controller incommunication with the sensors is configured to control flow of theblood fluid or lymph fluid through a lumen in response to the sensorhaving sensed the aptamer-based molecular beacon/cocaine complex in theblood fluid or lymph fluid. Once in the lumen of the device, theaptamer-based molecular beacon /cocaine complex binds to one or morebinding agents via the second structural element/peptide antigen of thebifunctional tag bound to an antibody in the treatment region of thedevice. In this manner, the cocaine is sequestered in the device andremoved from the circulation of the subject.

Each recited range includes all combinations and sub-combinations ofranges, as well as specific numerals contained therein.

All publications and patent applications cited in this specification areherein incorporated by reference to the extent not inconsistent with thedescription herein and for all purposes as if each individualpublication or patent application were specifically and individuallyindicated to be incorporated by reference for all purposes.

Those having ordinary skill in the art will recognize that the state ofthe art has progressed to the point where there is little distinctionleft between hardware and software implementations of aspects ofsystems; the use of hardware or software is generally (but not always,in that in certain contexts the choice between hardware and software canbecome significant) a design choice representing cost vs. efficiencytradeoffs. Those having ordinary skill in the art will recognize thatthere are various vehicles by which processes and/or systems and/orother technologies disclosed herein can be effected (e.g., hardware,software, and/or firmware), and that the preferred vehicle will varywith the context in which the processes and/or systems and/or othertechnologies are deployed. For example, if a surgeon determines thatspeed and accuracy are paramount, the surgeon may opt for a mainlyhardware and/or firmware vehicle; alternatively, if flexibility isparamount, the implementer may opt for a mainly software implementation;or, yet again alternatively, the implementer may opt for somecombination of hardware, software, and/or firmware. Hence, there areseveral possible vehicles by which the processes and/or devices and/orother technologies disclosed herein may be effected, none of which isinherently superior to the other in that any vehicle to be utilized is achoice dependent upon the context in which the vehicle will be deployedand the specific concerns (e.g., speed, flexibility, or predictability)of the implementer, any of which may vary. Those having ordinary skillin the art will recognize that optical aspects of implementations willtypically employ optically-oriented hardware, software, and or firmware.

In a general sense the various aspects disclosed herein which can beimplemented, individually and/or collectively, by a wide range ofhardware, software, firmware, or any combination thereof can be viewedas being composed of various types of “electrical circuitry.”Consequently, as used herein “electrical circuitry” includes, but is notlimited to, electrical circuitry having at least one discrete electricalcircuit, electrical circuitry having at least one integrated circuit,electrical circuitry having at least one application specific integratedcircuit, electrical circuitry forming a general purpose computing deviceconfigured by a computer program (e.g., a general purpose computerconfigured by a computer program which at least partially carries outprocesses and/or devices disclosed herein, or a microprocessorconfigured by a computer program which at least partially carries outprocesses and/or devices disclosed herein), electrical circuitry forminga memory device (e.g., forms of random access memory), and/or electricalcircuitry forming a communications device (e.g., a modem, communicationsswitch, or optical-electrical equipment). The subject matter disclosedherein may be implemented in an analog or digital fashion or somecombination thereof.

The herein described components (e.g., steps), devices, and objects andthe description accompanying them are used as examples for the sake ofconceptual clarity and that various configuration modifications usingthe disclosure provided herein are within the skill of those in the art.Consequently, as used herein, the specific examples set forth and theaccompanying description are intended to be representative of their moregeneral classes. In general, use of any specific example herein is alsointended to be representative of its class, and the non-inclusion ofsuch specific components (e.g., steps), devices, and objects hereinshould not be taken as indicating that limitation is desired.

With respect to the use of substantially any plural or singular termsherein, the reader can translate from the plural to the singular or fromthe singular to the plural as is appropriate to the context orapplication. The various singular/plural permutations are not expresslyset forth herein for sake of clarity.

The herein described subject matter sometimes illustrates differentcomponents contained within, or connected with, different othercomponents. It is to be understood that such depicted architectures aremerely examples, and that in fact many other architectures can beimplemented which achieve the same functionality. In a conceptual sense,any arrangement of components to achieve the same functionality iseffectively “associated” such that the desired functionality isachieved. Hence, any two components herein combined to achieve aparticular functionality can be seen as “associated with” each othersuch that the desired functionality is achieved, irrespective ofarchitectures or intermedial components. Likewise, any two components soassociated can also be viewed as being “operably connected,” or“operably coupled,” to each other to achieve the desired functionality,and any two components capable of being so associated can also be viewedas being “operably couplable,” to each other to achieve the desiredfunctionality. Specific examples of operably couplable include but arenot limited to physically mateable or physically interacting componentsor wirelessly interactable or wirelessly interacting components orlogically interacting or logically interactable components.

While particular aspects of the present subject matter described hereinhave been shown and described, changes and modifications may be madewithout departing from the subject matter described herein and itsbroader aspects and, therefore, the appended claims are to encompasswithin their scope all such changes and modifications as are within thetrue spirit and scope of the subject matter described herein.Furthermore, it is to be understood that the invention is defined by theappended claims. It will be understood that, in general, terms usedherein, and especially in the appended claims (e.g., bodies of theappended claims) are generally intended as “open” terms (e.g., the term“including” should be interpreted as “including but not limited to,” theterm “having” should be interpreted as “having at least,” the term“includes” should be interpreted as “includes but is not limited to,”etc.). It will be further understood that if a specific number of anintroduced claim recitation is intended, such an intent will beexplicitly recited in the claim, and in the absence of such recitationno such intent is present. For example, as an aid to understanding, thefollowing appended claims may contain usage of the introductory phrases“at least one” and “one or more” to introduce claim recitations.However, the use of such phrases should not be construed to imply thatthe introduction of a claim recitation by the indefinite articles “a” or“an” limits any particular claim containing such introduced claimrecitation to inventions containing only one such recitation, even whenthe same claim includes the introductory phrases “one or more” or “atleast one” and indefinite articles such as “a” or “an”; the same holdstrue for the use of definite articles used to introduce claimrecitations. In addition, even if a specific number of an introducedclaim recitation is explicitly recited, such recitation should typicallybe interpreted to mean at least the recited number (e.g., the barerecitation of “two recitations,” without other modifiers, typicallymeans at least two recitations, or two or more recitations).Furthermore, in those instances where a convention analogous to “atleast one of A, B, and C, etc.” is used, in general such a constructionis intended in the sense one having skill in the art would understandthe convention (e.g., “a system having at least one of A, B, and C”would include but not be limited to systems that have A alone, B alone,C alone, A and B together, A and C together, B and C together, or A, B,and C together, etc.). In those instances where a convention analogousto “at least one of A, B, or C, etc.” is used, in general such aconstruction is intended in the sense one having skill in the art wouldunderstand the convention (e.g., “a system having at least one of A, B,or C” would include but not be limited to systems that have A alone, Balone, C alone, A and B together, A and C together, B and C together, orA, B, and C together, etc.). Virtually any disjunctive word and/orphrase presenting two or more alternative terms, whether in thedescription, claims, or drawings, should be understood to contemplatethe possibilities of including one of the terms, either of the terms, orboth terms. For example, the phrase “A or B” will be understood toinclude the possibilities of “A” or “B” or “A and B.”

While various aspects and embodiments have been disclosed herein, otheraspects and embodiments will be apparent to those skilled in the art.The various aspects and embodiments disclosed herein are for purposes ofillustration and are not intended to be limiting, with the true scopeand spirit being indicated by the following claims.

1. An implantable device comprising: a body defining at least one lumen configured for fluid flow; at least one first reservoir disposed within the at least one lumen and configured to include one or more bifunctional tags, wherein the one or more bifunctional tags are configured to selectively bind to one or more target components in one or more of blood fluid or lymph fluid of a vertebrate subject; at least one treatment region disposed within the at least one lumen; and at least one second reservoir disposed in the at least one treatment region and configured to include one or more reactive components, wherein the one or more reactive components are configured to sequester the one or more bifunctional tags when bound to the one or more target components.
 2. The device of claim 1, wherein the one or more reactive components is configured to have an increased affinity for the one or more bifunctional tags bound to the one or more target components compared to an affinity for the one or more bifunctional tags unbound to the one or more reactive components.
 3. The device of claim 1, wherein the one or more reactive components is configured to modulate a physiological effect of the one or more target components.
 4. The device of claim 1, wherein the one or more bifunctional tags are configured to enter a circulatory system of the mammalian subject at a site different from a site of the one or more reactive components.
 5. The device of claim 1, wherein the one or more bifunctional tags includes one or more of a recognition element, recognition molecule, antibody, integrin, selectin, lectin, mimetic polymer, affibody, a label, or virus-like particle.
 6. The device of claim 5, wherein the label includes one or more of a QDOT, a nanoparticle, a fluorescent molecule, a magnetic particle, a contrast agent, or a radioisotope.
 7. The device of claim 5, wherein the one or more bifunctional tags includes one or more bifunctional antibodies.
 8. The device of claim 7, wherein the one or more bifunctional antibodies is configured to bind to one or more of the target component and the reactive component.
 9. The device of claim 1, wherein the one or more target components include one or more of circulating target cells or circulating target emboli.
 10. The device of claim 1, wherein the one or more target components includes one or more of tumor cells, emboli, misfolded proteins, aggregated proteins, antibodies, autoimmune antibodies, infectious agents, or infected cells.
 11. The device of claim 9, wherein the one or more target components include cancer cells, pre-cancer cells, autoimmune-related cells, B cells, T cells, phagocytes, platelets, lipoproteins, parasites, viruses, bacteria, fungi, or infected cells.
 12. The device of claim 1, further including two or more parallel lumen configured to receive the one or more target components.
 13. The device of claim 12, wherein a diameter of each of the two or more lumen is approximately less than two cell diameters.
 14. The device of claim 12, wherein a diameter of each of the two or more lumen is approximately less than 10 μm.
 15. (canceled)
 16. The device of claim 9, wherein the one or more reactive components is configured to produce necrosis or apoptosis in one or more target cells.
 17. The device of claim 1, wherein the one or more reactive components is configured to alter, arrest, or destroy the one or more target components.
 18. The device of claim 1, wherein the one or more reactive components is configured to be placed relative to a tumor or an organ in the mammalian subject.
 19. The device of claim 1, wherein the one or more reactive components includes one or more of an adhesion molecule, antibody, binding mimetic, polymer, lectin, integrin, or selectin.
 20. The device of claim 1, wherein the one or more reactive components include one or more of a denaturing agent, degradative agent, or binding agent.
 21. The device of claim 20, wherein the one or more reactive components include a cytotoxic agent, a cytostatic agent, a programmed cell death-inducing agent, a chemotherapeutic agent, or an antibody-toxin agent.
 22. The device of claim 20, wherein the one or more binding agents include one or more of antibodies, receptors, or cognates configured to bind to one or more target components.
 23. The device of claim 20, wherein the one or more binding agents include one or more of lectin, binding protein, catalytic antibody, catalytic aptamer, protease conjugate, or photoactivatable conjugate.
 24. The device of claim 1, further including one or more energy sources configured to supply energy to the at least one treatment region.
 25. The device of claim 1, further including one or more sensor configured to measure a physiological condition proximate to the device.
 26. The device of claim 25, wherein the one or more sensor is configured to detect one or more of unbound bifunctional tags or bifunctional tags bound to target components in the one or more of blood fluid or lymph fluid of the vertebrate subject.
 27. The device of claim 25, wherein the one or more sensor is configured to detect sequestration by the at least one reactive component.
 28. The device of claim 25, further including a transmitter to report data from the one or more sensor.
 29. The device of claim 25, wherein the one or more sensor is configured to report to an outside source or to a computing device.
 30. The device of claim 25, wherein the one or more sensor is configured to function in, or proximal to, the one or more blood vessel or lymph vessel.
 31. The device of claim 25, wherein the one or more sensor is external to the at least one lumen.
 32. The device of claim 25, wherein the one or more sensor is internal to the at least one lumen.
 33. The device of claim 25, further including at least one controller in communication with the one or more sensor.
 34. (canceled)
 35. The device of claim 33, further including at least one controllable flow barrier to fluid flow into the at least one lumen.
 36. (canceled)
 37. The device of claim 35, wherein the at least one controller in communication with the one or more sensor is configured to control the at least one controllable flow barrier to the at least one lumen.
 38. The device of claim 33, wherein the one or more sensor is configured to detect the one or more target components and configured to communicate with the at least one controller to release the one or more bifunctional tags in response to the one or more detected target components.
 39. The device of claim 33, wherein the one or more sensor is configured to detect the one or more bifunctional tags complexed with the one or more target components and is configured to communicate with the controller to activate the one or more reactive components in response to the complex of the one or more bifunctional tags to the one or more target components.
 40. The device of claim 37, wherein the one or more sensor is configured to detect the one or more bifunctional tags complexed with the one or more target components and is configured to communicate with the controller to divert flow of the one or more of blood fluid or lymph fluid to the at least one lumen of the device.
 41. (canceled)
 42. The device of claim 1, wherein the one or more reactive components are configured to attach to the at least one lumen.
 43. The device of claim 33, wherein the one or more sensor and the at least one controller are configured to achieve a target level of the one or more target components in the vertebrate subject.
 44. The device of claim 43, wherein the one or more sensor and the at least one controller are configured to control the at least one controllable flow barrier, to activate the one or more reactive components, to release the one or more bifunctional tags, or to activate one or more energy sources.
 45. The device of claim 43, wherein the one or more sensor and the at least one controller are configured to control levels of the detected one or more target components to limit a deviation from the target level.
 46. The device of claim 45, wherein the deviation is determined by a weighted least squares fit.
 47. The device of claim 43, wherein the target level includes a desired concentration of the one or more target components in the one or more of blood fluid or lymph fluid.
 48. The device of claim 43, wherein the target level includes a desired range of concentrations of the one or more target components in the one or more of blood fluid or lymph fluid.
 49. The device of claim 43, wherein the target level includes a desired ratio of concentrations of two or more target components in the one or more of blood fluid or lymph fluid.
 50. The device of claim 43, wherein the target level includes a desired ratio of levels of two or more target components in the one or more of blood fluid or lymph fluid.
 51. The device of claim 33, wherein the at least one first reservoir configured to provide one or more bifunctional tags is responsive to the controller.
 52. The device of claim 33, wherein the at least one second reservoir configured to provide one or more reactive components is responsive to the controller.
 53. The device of claim 25, wherein the one or more sensor includes a biosensor, chemical sensor, physical sensor, or optical sensor.
 54. The device of claim 53, wherein the one or more sensor includes one or more target recognition elements. 55.-57. (canceled)
 58. The device of claim 25, wherein the one or more sensor is configured to target the device to a site having an elevated level of the one or more target components. 59.-65. (canceled)
 66. The device of claim 20, wherein the one or more second reservoirs include a matrix configured to present one or more reactive components. 67.-79. (canceled)
 80. The device of claim 24, wherein the energy source is coupled to one or more sensor configured to selectively direct energy to the target component. 81.-84. (canceled)
 85. The device of claim 1, wherein the one or more second reservoirs include a source for producing the one or more reactive components, the source including at least one reservoir and at least one producer. 86.-447. (canceled) 